LOCUS       piG17-1-003_Plux        4139 bp    DNA     circular UNA 01-NOV-2017
DEFINITION  Concatenation of 4 sequences.
ACCESSION   urn.local...4d-88wu309
VERSION     urn.local...4d-88wu309
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          <1..2827
                     /organism
                     /label="source"
     terminator      7..101
                     /locus_tag="/note=""transcription terminator from phage
                     lambda"
                     /product
                     /label="lambda t0 terminator"
     CDS             233..1048
                     /locus_tag="/note=""confers resistance to kanamycin in
                     bacteria or G418 (Geneticin®) in eukaryotes"
                     /product="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
                     /label="KanR"
     oriT            1204..1312
                     /locus_tag="/note=""incP origin of transfer"
                     /product
                     /label="oriT"
     rep_origin      1348..1936
                     /locus_tag="/note=""high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of rep lication"
                     /product
                     /label="ori"
     misc_feature    2122..2262
                     /locus_tag="/product="
                     /label="bom"
     CDS             complement(2364..2555)
                     /locus_tag="/product=""Rop protein, which maintains
                     plasmids at low copy number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
                     /label="rop"
     terminator      2735..2821
                     /locus_tag="/note=""transcription terminator T1 from the
                     E. coli rrnB ge ne"
                     /product
                     /label="rrnB T1 terminator"
     misc_feature    2782
                     /locus_tag="/product="
                     /label="A"
     DNA_probe_bind  2788..2867
                     /created_by="cherasv"
                     /Sequence="CCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTGATGCCTGAATTCG
                     CGGCCGCTTCTAGAGACCTGTAGGATCGTACAG"
                     /Mismatches=0
                     /annotation_group="8ee9ff6a-6df3-498a-b8cd-cb9e6dbdd412"
                     /label="oiG17-00_LCR_bridge_Backbone_Plux"
     misc_feature    2828..2910
                     /Source="referenceToDocument: urn:local:.:4h-88wu309"
                     /label="ciG17-006_Plux (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    2828..2849
                     /created_by="cherasv"
                     /modified_by="cherasv"
                     /label="BioBrick Prefix"
     promoter        2850..2910
                     /created_by="cherasv"
                     /label="Plux "
     DNA_probe_bind  2871..2950
                     /annotation_group="6a42bf05-2571-45f1-8f10-00cf262c9ca2"
                     /Mismatches=0
                     /label="oiG17-00_LCR_bridge_Plux_RBS_azurin"
     source          <2911..>3351
                     /organism="Pseudomonas aeruginosa"
                     /mol_type="genomic DNA"
                     /strain="CIT135"
                     /db_xref="taxon:287"
                     /label="source Pseudomonas aeruginosa"
     misc_feature    2911..2952
                     /Salis_TIR=32320
                     /created_by="cherasv"
                     /modified_by="cherasv"
                     /label="gcggt+RBS"
     CDS             2953..3342
                     /gene="azu"
                     /note="precursor"
                     /codon_start=1
                     /transl_table=11
                     /product="azurin"
                     /protein_id="AAA25730.1"
                     /translation="MLRKLAAVSLLSLLSAPLLAAECSVDIQGNDQMQFNTNAITVDK
                     SCKQFTVNLSHPGNLPKNVMGHNWVLSTAADMQGVVTDGMASGLDKDYLKPDDSRVIA
                     HTKLIGSGEKDSVTFDVSKLKEGEQYMFFCTFPGHSALMKGTLTLK"
                     /label="azu CDS"
     source          <2953..>3342
                     /organism="Pseudomonas aeruginosa"
                     /mol_type="genomic DNA"
                     /strain="CIT135"
                     /db_xref="taxon:287"
                     /label="source Pseudomonas aeruginosa"
     Optimized_Codon 2956..2958
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /modified_by="cherasv"
                     /label="GCG"
     Optimized_Codon 2959..2961
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 2965..2967
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 2968..2970
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3007..3009
                     /Relative_Adaptiveness="1.0"
                     /Original="AAT"
                     /label="AAC"
     Optimized_Codon 3010..3012
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3019..3021
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3025..3027
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3028..3030
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3034..3036
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3046..3048
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3055..3057
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3061..3063
                     /Relative_Adaptiveness="1.0"
                     /Original="CCC"
                     /label="CCG"
     Optimized_Codon 3064..3066
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3076..3078
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3082..3084
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3088..3090
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3100..3102
                     /Relative_Adaptiveness="1.0"
                     /Original="GTA"
                     /label="GTT"
     Optimized_Codon 3106..3108
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3112..3114
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3115..3117
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3127..3129
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3130..3132
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3133..3135
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3142..3144
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3148..3150
                     /Relative_Adaptiveness="1.0"
                     /Original="GCT"
                     /label="GCG"
     Optimized_Codon 3151..3153
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3154..3156
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3163..3165
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3166..3168
                     /Relative_Adaptiveness="1.0"
                     /Original="GAT"
                     /label="GAC"
     Optimized_Codon 3175..3177
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3178..3180
                     /Relative_Adaptiveness="1.0"
                     /Original="CCC"
                     /label="CCG"
     Optimized_Codon 3187..3189
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3190..3192
                     /Relative_Adaptiveness="1.0"
                     /Original="CGC"
                     /label="CGT"
     Optimized_Codon 3193..3195
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3199..3201
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3208..3210
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3217..3219
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3220..3222
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 3223..3225
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3226..3228
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 3229..3231
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3235..3237
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 3238..3240
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3250..3252
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3253..3255
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3256..3258
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3262..3264
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3268..3270
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3271..3273
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 3301..3303
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3307..3309
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     DNA_probe_bind  3312..3391
                     /annotation_group="2c9c76ca-5fe5-4a5e-a1d3-3fc3c3cdae26"
                     /Mismatches=0
                     /label="oiG17-00_bridge_LCR_azurin_gfp"
     Optimized_Codon 3319..3321
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3322..3324
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3337..3339
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3340..3342
                     /Relative_Adaptiveness="1.0"
                     /Original="TGA"
                     /label="TAA"
     misc_feature    <3343..3351
                     /created_by="cherasv"
                     /modified_by="cherasv"
                     /label="SacI_GTC_ext"
     misc_feature    3343..3348
                     /Recognition_pattern="GAGCT^C"
                     /annotation_group="GAGCTC"
                     /label="SacI, SstI"
                     /note="Geneious type: restriction site"
     misc_feature    3344..3347
                     /Recognition_pattern="AG^CT"
                     /annotation_group="AGCT"
                     /label="AluI"
                     /note="Geneious type: restriction site"
     misc_feature    3352..3357
                     /Recognition_pattern="G^TCGAC"
                     /label="SalI"
                     /note="Geneious type: restriction site"
     RBS             3358..3392
                     /locus_tag="/product="
                     /label="strong RBS"
     source          <3358..>4118
                     /organism
                     /label="source"
     CDS             3393..4118
                     /locus_tag="/note=""sfGFP"
                     /product="/translation=""MSKGEELFTGVVPILVELDGDVNGHKFSVRGEG
                     EGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNG
                     IKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYKRP"
                     /label="sfGFP"
     DNA_probe_bind  4100..40
                     /Sequence="CTACAAAAGGCCTTAATAATACTAGTAGCGGCCGCTGCAGGTCTTGG
                     ACTCCTGTTGATAGATCCAGTAATGACCTCAGA"
                     /created_by="cherasv"
                     /label="oiG17-00_LCR_bridge_gfp_SF_backbone"
     misc_feature    4119..4139
                     /created_by="cherasv"
                     /label="BioBrickSuffix"
ORIGIN      
        1 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
       61 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt
      121 acgatttaaa tttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
      181 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
      241 tattcagcgt gaaacgagct gtagccgtcc gcgtctgaac agcaacatgg atgcggatct
      301 gtatggctat aaatgggcgc gtgataacgt gggtcagagc ggcgcgacca tttatcgtct
      361 gtatggcaaa ccggatgcgc cggaactgtt tctgaaacat ggcaaaggca gcgtggcgaa
      421 cgatgtgacc gatgaaatgg tgcgtctgaa ctggctgacc gaatttatgc cgctgccgac
      481 cattaaacat tttattcgca ccccggatga tgcgtggctg ctgaccaccg cgattccggg
      541 caaaaccgcg tttcaggtgc tggaagaata tccggatagc ggcgaaaaca ttgtggatgc
      601 gctggccgtg tttctgcgtc gtctgcatag cattccggtg tgcaactgcc cgtttaacag
      661 cgatcgtgtg tttcgtctgg cccaggcgca gagccgtatg aacaacggcc tggtggatgc
      721 gagcgatttt gatgatgaac gtaacggctg gccggtggaa caggtgtgga aagaaatgca
      781 taaactgctg ccgtttagcc cggatagcgt ggtgacccac ggcgatttta gcctggataa
      841 cctgattttc gatgaaggca aactgattgg ctgcattgat gtgggccgtg tgggcattgc
      901 ggatcgttat caggatctgg ccattctgtg gaactgcctg ggcgaattta gcccgagcct
      961 gcaaaaacgt ctgtttcaga aatatggcat tgataatccg gatatgaaca aactgcaatt
     1021 tcatctgatg ctggatgaat ttttctaata attaattgga caagggtcct tttccgctgc
     1081 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     1141 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     1201 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     1261 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg
     1321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2101 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2161 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2221 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2281 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2341 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2401 cagctgcggt aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca
     2461 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2521 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     2581 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     2641 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     2701 cggcgggggc gcgcccagct gtctagggcg gcggatttgt cctactcagg agagcgttca
     2761 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     2821 gatgcctgaa ttcgcggccg cttctagaga cctgtaggat cgtacaggtt tacgcaagaa
     2881 aatggtttgt tatagtcgaa taaaccatgg gcggtcctta tatcatcatc aattcactaa
     2941 gggggttaca ttatggcgga atgctctgtt gacatccagg gtaacgacca gatgcagttc
     3001 aacaccaacg cgatcaccgt tgacaaatct tgcaaacagt tcaccgttaa cctgtctcac
     3061 ccgggtaacc tgccgaaaaa cgttatgggt cacaactggg ttctgtctac cgcggcggac
     3121 atgcagggtg ttgttaccga cggtatggcg tctggtctgg acaaagacta cctgaaaccg
     3181 gacgactctc gtgttatcgc gcacaccaaa ctgatcggtt ctggtgaaaa agactctgtt
     3241 accttcgacg tttctaaact gaaagaaggt gaacagtaca tgttcttctg caccttcccg
     3301 ggtcactctg cgctgatgaa aggtaccctg accctgaaat aagagctcgt cgtcgacgat
     3361 aaatgcttca ataatatgag gaggtgatct agatgagcaa aggagaagaa cttttcactg
     3421 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtcc
     3481 gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt atttgcacta
     3541 ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat ggtgttcaat
     3601 gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt gccatgcccg
     3661 aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac aagacgcgtg
     3721 ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag ggtattgatt
     3781 ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac tcacacaatg
     3841 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca
     3901 acgttgaaga tggttccgtt caactagcag accattatca acaaaatact ccaattggcg
     3961 atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc ctttcgaaag
     4021 atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta
     4081 cacatggcat ggatgagctc tacaaaaggc cttaataata ctagtagcgg ccgctgcag
//
