LOCUS       sequence-388753-        3632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of J23103 and B0034m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3632)
  AUTHORS   Keating KW, Young EM
  TITLE     Systematic Part Transfer by Extending a Modular Toolkit to Diverse 
            Bacteria.
  JOURNAL   ACS Synth Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104.
  PUBMED    37294017
REFERENCE   2  (bases 1 to 3632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104."
COMMENT     Sequence Label: pBBR1_DVK_AE(J23103-B0034m-E0040m-B0015)
FEATURES             Location/Qualifiers
     source          1..3632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=BBa_J23112
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0"
     promoter        5..39
                     /label=BBa_J23103
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     RBS             48..59
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             69..782
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(99..127)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     722..742
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      801..872
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      888..915
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      1063..1834
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1835..2497
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     oriT            2508..2616
                     /note="incP origin of transfer"
     primer_bind     complement(2706..2725)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2717..3532
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2717..3532
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2790..2809)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggagctgata gctagctcag tcctagggat tatgctagct actagagaaa gaggagaaat
       61 actaaatgat gcgtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      121 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      181 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
      241 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
      301 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
      361 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
      421 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
      481 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
      541 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
      601 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      661 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagatcaca
      721 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
      781 aataataaag gtccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      841 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
      901 gtgggccttt ctgcgtttat agcttatgtc ttcagcagta ggacaaatcc gccgccctag
      961 acagctgggc gcgccccccc tacgggcttg ctctccgggc ttcgccctgc gcggtcgctg
     1021 cgctcccttg ccagcccgtg gatatgtgga cgatggccgc gagcggccac cggctggctc
     1081 gcttcgctcg gcccgtggac aaccctgctg gacaagctga tggacaggct gcgcctgccc
     1141 acgagcttga ccacagggat tgcccaccgg ctacccagcc ttcgaccaca tacccaccgg
     1201 ctccaactgc gcggcctgcg gccttgcccc atcaattttt ttaattttct ctggggaaaa
     1261 gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg acaccaagtg gaaggcgggt
     1321 caaggctcgc gcagcgaccg cgcagcggct tggccttgac gcgcctggaa cgacccaagc
     1381 ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc ctgccccccg agcctcacgg
     1441 cggcgagtgc gggggttcca agggggcagc gccaccttgg gcaaggccga aggccgcgca
     1501 gtcgatcaac aagccccgga ggggccactt tttgccggag ggggagccgc gccgaaggcg
     1561 tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa gaactagata tagggcgaaa
     1621 tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt acgcaacagc tcattgcggc
     1681 accccccgca atagctcatt gcgtaggtta aagaaaatct gtaattgact gccactttta
     1741 cgcaacgcat aattgttgtc gcgctgccga aaagttgcag ctgattgcgc atggtgccgc
     1801 aaccgtgcgg cacccctacc gcatggagat aagcatggcc acgcagtcca gagaaatcgg
     1861 cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg gaacgcaaag cgcatgaggc
     1921 gtgggccggg cttattgcga ggaaacccac ggcggcaatg ctgctgcatc acctcgtggc
     1981 gcagatgggc caccagaacg ccgtggtggt cagccagaaa acactttcca agctcatcgg
     2041 acgttctttg cggacggtcc aatacgcagt caaggacttg gtggccgagc gctggatctc
     2101 cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac gtggtcaatg accgcgtggc
     2161 gtggggccag ccccgcgacc agttgcgcct gtcggtgttc agtgccgccg tggtggttga
     2221 tcacgacgac caggacgaat cgctgttggg gcatggcgac ctgcgccgca tcccgaccct
     2281 gtatccgggc gagcagcaac taccgaccgg ccccggcgag gagccgccca gccagcccgg
     2341 cattccgggc atggaaccag acctgccagc cttgaccgaa acggaggaat gggaacggcg
     2401 cgggcagcag cgcctgccga tgcccgatga gccgtgtttt ctggacgatg gcgagccgtt
     2461 ggagccgccg acacgggtaa cgctgccgcg ccggtagggc cggcctacgg ccagcctcgc
     2521 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     2581 ccgctcgcgg gtgggcctac ttcacctatc ctgcccttgt gtctcaaaat ctctgatgta
     2641 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2701 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcca ggccgcgatt
     2761 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2821 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2881 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2941 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3001 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     3061 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3121 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3181 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3241 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     3301 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3361 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3421 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3481 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aaggtaactg
     3541 tcagaccaag tttactcata tatactttag attgattttt cgttccactg agcgtcagac
     3601 cccagaactc tgtcagaccg agtggaagac at
//
