LOCUS       piG17-1-010             3710 bp    DNA     circular UNA 01-NOV-2017
DEFINITION  Concatenation of 5 sequences.
ACCESSION   urn.local...et-88wu36t
VERSION     urn.local...et-88wu36t
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          <1..2827
                     /organism
                     /label="source"
     misc_feature    1..2827
                     /Source="referenceToDocument: urn:local:.:er-88wu36t"
                     /label="piG17-001_actuator_backbone (concatenated sequence
                     1)"
                     /note="Geneious type: Concatenated sequence"
     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  2828..2910
                     /Sequence="GAATTCGCGGCCGCTTCTAGAGACCTGTAGGATCGTACAGGTTTACG
                     CAAGAAAATGGTTTGTTATAGTCGAATAAACCATGG"
                     /created_by="irma"
                     /label="ciG17-006"
     misc_feature    2828..2910
                     /Source="referenceToDocument: urn:local:.:es-88wu36t"
                     /label="ciG17-006 (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     RBS             2911..2937
                     /label="Protein E max RBS"
     misc_feature    2911..3216
                     /Source="referenceToDocument: urn:local:.:ev-88wu36t"
                     /label="pIW91_pIW281_rhaRS.proteinE PCR Product without SF
                     (concatenated sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     CDS             2938..>3210
                     /codon_start=1
                     /translation="MVRWTLWDTLAFLLLLSLLLPSLLIMFIPSTFKRPVSSWKALNL
                     RKTLLMASSVRLKPLNCSRLPCVYAQETLTFLLTQKKTCVKNYVRKE"
                     /label="proteinE phiX174"
     source          <2938..>3210
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     misc_feature    complement(3211..3213)
                     /label="rev compl stop codon"
     misc_feature    complement(3214..3216)
                     /label="rev compl stop codon"
     misc_feature    3217..3263
                     /Source="referenceToDocument: urn:local:.:eq-88wu36t"
                     /label="ciG17-016 (concatenated sequence 4)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    complement(3217..3222)
                     /created_by="irma"
                     /modified_by="irma"
                     /label="1-RS2 (SacI)"
     promoter        3223..3257
                     /created_by="lidavadakumchery"
                     /label="BBa_J23118"
     misc_feature    complement(3258..3263)
                     /created_by="irma"
                     /label="RS3 (SalI)"
     source          <3264..>3710
                     /organism="Pseudomonas aeruginosa"
                     /mol_type="genomic DNA"
                     /strain="CIT135"
                     /db_xref="taxon:287"
                     /label="source Pseudomonas aeruginosa"
     misc_feature    3264..3710
                     /Source="referenceToDocument: urn:local:.:eu-88wu36t"
                     /label="ciG17-009_RBS_azurin_SF_no_signal_peptide
                     (concatenated sequence 5)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    3264..3293
                     /TIR_Salis=3625576
                     /created_by="cherasv"
                     /label="Synthetic_RBS"
     CDS             3294..3683
                     /gene="azu"
                     /note="precursor"
                     /codon_start=1
                     /transl_table=11
                     /product="azurin"
                     /protein_id="AAA25730.1"
                     /translation="MLRKLAAVSLLSLLSAPLLAAECSVDIQGNDQMQFNTNAITVDK
                     SCKQFTVNLSHPGNLPKNVMGHNWVLSTAADMQGVVTDGMASGLDKDYLKPDDSRVIA
                     HTKLIGSGEKDSVTFDVSKLKEGEQYMFFCTFPGHSALMKGTLTLK"
                     /label="azu CDS"
     source          <3294..>3683
                     /organism="Pseudomonas aeruginosa"
                     /mol_type="genomic DNA"
                     /strain="CIT135"
                     /db_xref="taxon:287"
                     /label="source Pseudomonas aeruginosa"
     Optimized_Codon 3297..3299
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3300..3302
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 3306..3308
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 3309..3311
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3348..3350
                     /Relative_Adaptiveness="1.0"
                     /Original="AAT"
                     /label="AAC"
     Optimized_Codon 3351..3353
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3360..3362
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3366..3368
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3369..3371
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3375..3377
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3387..3389
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3396..3398
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3402..3404
                     /Relative_Adaptiveness="1.0"
                     /Original="CCC"
                     /label="CCG"
     Optimized_Codon 3405..3407
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3417..3419
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3423..3425
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3429..3431
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3441..3443
                     /Relative_Adaptiveness="1.0"
                     /Original="GTA"
                     /label="GTT"
     Optimized_Codon 3447..3449
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3453..3455
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3456..3458
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3468..3470
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3471..3473
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3474..3476
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3483..3485
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3489..3491
                     /Relative_Adaptiveness="1.0"
                     /Original="GCT"
                     /label="GCG"
     Optimized_Codon 3492..3494
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3495..3497
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3504..3506
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3507..3509
                     /Relative_Adaptiveness="1.0"
                     /Original="GAT"
                     /label="GAC"
     Optimized_Codon 3516..3518
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3519..3521
                     /Relative_Adaptiveness="1.0"
                     /Original="CCC"
                     /label="CCG"
     Optimized_Codon 3528..3530
                     /Relative_Adaptiveness="1.0"
                     /Original="AGC"
                     /label="TCT"
     Optimized_Codon 3531..3533
                     /Relative_Adaptiveness="1.0"
                     /Original="CGC"
                     /label="CGT"
     Optimized_Codon 3534..3536
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3540..3542
                     /Relative_Adaptiveness="1.0"
                     /Original="GCC"
                     /label="GCG"
     Optimized_Codon 3549..3551
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3558..3560
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3561..3563
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 3564..3566
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3567..3569
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 3570..3572
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3576..3578
                     /Relative_Adaptiveness="1.0"
                     /Original="TCG"
                     /label="TCT"
     Optimized_Codon 3579..3581
                     /Relative_Adaptiveness="1.0"
                     /Original="GTG"
                     /label="GTT"
     Optimized_Codon 3591..3593
                     /Relative_Adaptiveness="1.0"
                     /Original="GTC"
                     /label="GTT"
     Optimized_Codon 3594..3596
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3597..3599
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3603..3605
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3609..3611
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3612..3614
                     /Relative_Adaptiveness="1.0"
                     /Original="GAG"
                     /label="GAA"
     Optimized_Codon 3642..3644
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3648..3650
                     /Relative_Adaptiveness="1.0"
                     /Original="TCC"
                     /label="TCT"
     Optimized_Codon 3660..3662
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3663..3665
                     /Relative_Adaptiveness="1.0"
                     /Original="GGC"
                     /label="GGT"
     Optimized_Codon 3678..3680
                     /Relative_Adaptiveness="1.0"
                     /Original="AAG"
                     /label="AAA"
     Optimized_Codon 3681..3683
                     /Relative_Adaptiveness="1.0"
                     /Original="TGA"
                     /label="TAA"
     misc_feature    3684..>3710
                     /created_by="cherasv"
                     /modified_by="cherasv"
                     /label="Suffix+TATTTA"
     misc_feature    3692..3699
                     /Recognition_pattern="GC^GGCCGC"
                     /annotation_group="GCGGCCGC"
                     /label="NotI"
                     /note="Geneious type: restriction site"
     misc_feature    3694..3697
                     /Recognition_pattern="GG^CC"
                     /annotation_group="GGCC"
                     /label="HaeIII"
                     /note="Geneious type: restriction site"
     misc_feature    3699..3704
                     /Recognition_pattern="CTGCA^G"
                     /annotation_group="CTGCAG"
                     /label="PstI"
                     /note="Geneious type: restriction site"
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 cacaataaga atttaaggag gtaaaatatg
     2941 gtacgctgga ctttgtggga taccctcgct ttcctgctcc tgttgagttt attgctgccg
     3001 tcattgctta ttatgttcat cccgtcaaca ttcaaacggc ctgtctcatc atggaaggcg
     3061 ctgaatttac ggaaaacatt attaatggcg tcgagcgtcc ggttaaagcc gctgaattgt
     3121 tcgcgtttac cttgcgtgta cgcgcaggaa acactgacgt tcttactgac gcagaagaaa
     3181 acgtgcgtca aaaattacgt gcggaaggag taataagagc tcttgacggc tagctcagtc
     3241 ctaggtattg tgctagcgtc gacaacaccg acatacaaat aaggaggtta aacatggcgg
     3301 aatgctctgt tgacatccag ggtaacgacc agatgcagtt caacaccaac gcgatcaccg
     3361 ttgacaaatc ttgcaaacag ttcaccgtta acctgtctca cccgggtaac ctgccgaaaa
     3421 acgttatggg tcacaactgg gttctgtcta ccgcggcgga catgcagggt gttgttaccg
     3481 acggtatggc gtctggtctg gacaaagact acctgaaacc ggacgactct cgtgttatcg
     3541 cgcacaccaa actgatcggt tctggtgaaa aagactctgt taccttcgac gtttctaaac
     3601 tgaaagaagg tgaacagtac atgttcttct gcaccttccc gggtcactct gcgctgatga
     3661 aaggtaccct gaccctgaaa taatactagt agcggccgct gcagtattta
//
