LOCUS       piG17-1-012b            4428 bp    DNA     circular UNA 14-OCT-2017
DEFINITION  Gibson Assembly ligation product.
ACCESSION   urn.local...97-85n9etc
VERSION     urn.local...97-85n9etc
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          <1..2827
                     /organism
     misc_feature    1..3802
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="Digestion of piG17-1-008b extraction
                     (concatenated sequence 1)"
     misc_feature    1..2827
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="piG17-001_actuator_backbone (concatenated
                     sequence 1)"
     primer_bind     1..25
                     /standard_name="oiG17-043"
     misc_feature    complement(1..25)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-51"
     terminator      7..101
                     /locus_tag="/note=""transcription terminator from phage
                     lambda"
                     /standard_name="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"
                     /standard_name="KanR"
     primer_bind     1079..1148
                     /standard_name="oiG17-013"
     misc_feature    complement(1091..1108)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-012"
     primer_bind     1109..1130
                     /standard_name="oiG17-011"
     oriT            1204..1312
                     /locus_tag="/note=""incP origin of transfer"
                     /standard_name="oriT"
     rep_origin      1348..1936
     misc_feature    2122..2262
                     /locus_tag="/product="
                     /standard_name="bom"
     CDS             complement(2364..2555)
                     /locus_tag="/product=""Rop protein, which maintains
                     plasmids at low copy number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
                     /standard_name="rop"
     primer_bind     2733..2752
                     /standard_name="oDG127"
     terminator      2735..2821
                     /locus_tag="/note=""transcription terminator T1 from the
                     E. coli rrnB ge ne"
                     /standard_name="rrnB T1 terminator"
     misc_feature    2782
                     /locus_tag="/product="
                     /standard_name="A"
     misc_feature    complement(2804..2827)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-008"
     primer_bind     2804..2827
                     /standard_name="oiG17-50"
     regulatory      2828..2849
                     /standard_name="Prefix"
     misc_feature    complement(2834..2849)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-67"
     regulatory      2850..2866
                     /standard_name="lldR binding site O1"
     regulatory      2867..2883
                     /standard_name="lldR binding site O1"
     promoter        2920..2974
                     /standard_name="BBa_R0062"
     primer_bind     2920..2941
                     /standard_name="oiG17-66"
     regulatory      2938..2944
                     /standard_name="-35"
     regulatory      2961..2966
                     /standard_name="-10"
     regulatory      2970
                     /standard_name
     regulatory      2994..3010
                     /standard_name="lldR binding site O2"
     regulatory      3011..3027
                     /standard_name="lldR binding site O2"
     misc_feature    3028..3033
                     /standard_name="RS4 (HindIII)"
     misc_feature    complement(3034..3039)
                     /standard_name="RS3 (SalI)"
     primer_bind     3034..3069
                     /standard_name="oiG17-003"
     primer_bind     3034..3053
                     /standard_name="oiG17-52"
     RBS             3040..3074
                     /locus_tag="/product="
                     /standard_name="strong RBS"
     source          <3040..>3800
                     /organism
     primer_bind     3040..3069
                     /standard_name="oiG17-001"
     CDS             3075..3800
                     /locus_tag="/note=""sfGFP"
                     /product="/translation=""MSKGEELFTGVVPILVELDGDVNGHKFSVRGEG
                     EGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNG
                     IKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYKRP"
                     /standard_name="sfGFP"
     misc_feature    complement(3779..3800)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-75"
     misc_feature    3782..3802
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
     misc_feature    3782..4428
                     /note="Geneious type: Concatenated sequence"
                     /standard_name="ciG17-005 PCR Product (concatenated
                     sequence 2)"
     misc_feature    3801..>3802
                     /standard_name="BioBrick Suffix"
     primer_bind     3803..3822
                     /standard_name="oiG17-72"
     CDS             3829..4416
                     /standard_name="LuxI"
     primer_bind     3829..3851
                     /standard_name="oiG17-63"
     misc_feature    complement(4410..4428)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="oiG17-53"
     misc_feature    4414..4428
                     /note="Geneious type: ligation"
                     /standard_name="Ligation"
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 attggcccta ccaattaatt ggccctacca
     2881 attcttcgct tatctgacct ctggttcaca atttcccaaa cctgtaggat cgtacaggtt
     2941 tacgcaagaa aatggtttgt tatagtcgaa taaatcatta tccctacaca acaaattggc
     3001 agtgccactt aattggcagt gccacttaag cttgtcgacg ataaatgctt caataatatg
     3061 aggaggtgat ctagatgagc aaaggagaag aacttttcac tggagttgtc ccaattcttg
     3121 ttgaattaga tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg
     3181 atgctacaaa cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc
     3241 cgtggccaac acttgtcact actctgacct atggtgttca atgcttttcc cgttatccgg
     3301 atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac
     3361 gcactatatc tttcaaagat gacgggacct acaagacgcg tgctgaagtc aagtttgaag
     3421 gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa gatggaaaca
     3481 ttcttggaca caaactcgag tacaacttta actcacacaa tgtatacatc acggcagaca
     3541 aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacgttgaa gatggttccg
     3601 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     3661 cagacaacca ttacctgtcg acacaatctg tcctttcgaa agatcccaac gaaaagcgtg
     3721 accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc
     3781 tctacaaaag gccttaataa taggggtaac ccgtaggtag gggggaaaat gaccatcatg
     3841 atcaaaaaat ctgacttcct ggcgatcccg tctgaagaat acaaaggtat cctgtctctg
     3901 cgttaccagg ttttcaaaca gcgtctggaa tgggacctgg ttgttgaaaa caacctggaa
     3961 tctgacgaat acgacaactc taacgcggaa tacatctacg cgtgcgacga caccgaaaac
     4021 gtttctggtt gctggcgtct gctgccgacc accggtgact acatgctgaa atctgttttc
     4081 ccggaactgc tgggtcagca gtctgcgccg aaagacccga acatcgttga actgtctcgt
     4141 ttcgcggttg gtaaaaactc ttctaaaatc aacaactctg cgtctgaaat caccatgaaa
     4201 ctgttcgaag cgatctacaa acacgcggtt tctcagggta tcaccgaata cgttaccgtt
     4261 acctctaccg cgatcgaacg tttcctgaaa cgtatcaaag ttccgtgcca ccgtatcggt
     4321 gacaaagaaa tccacgttct gggtgacacc aaatctgttg ttctgtctat gccgatcaac
     4381 gaacagttca aaaaagcggt tctgaactac tagtagcggc cgctgcag
//
