LOCUS       sequence-388754-        3631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of J23103 and B0033m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3631)
  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 3631)
  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-B0033m-E0040m-B0015)
FEATURES             Location/Qualifiers
     source          1..3631
                     /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"
     CDS             68..781
                     /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(98..126)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     721..741
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      800..871
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      887..914
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      1062..1833
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1834..2496
                     /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            2507..2615
                     /note="incP origin of transfer"
     primer_bind     complement(2705..2724)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2716..3531
                     /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             2716..3531
                     /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(2789..2808)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggagctgata gctagctcag tcctagggat tatgctagct actagagtca cacaggacta
       61 ctaaatgatg cgtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      121 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      181 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      241 aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat
      301 gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
      361 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
      421 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
      481 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
      541 gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
      601 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
      661 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagatcacat
      721 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
      781 ataataaagg tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      841 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
      901 tgggcctttc tgcgtttata gcttatgtct tcagcagtag gacaaatccg ccgccctaga
      961 cagctgggcg cgccccccct acgggcttgc tctccgggct tcgccctgcg cggtcgctgc
     1021 gctcccttgc cagcccgtgg atatgtggac gatggccgcg agcggccacc ggctggctcg
     1081 cttcgctcgg cccgtggaca accctgctgg acaagctgat ggacaggctg cgcctgccca
     1141 cgagcttgac cacagggatt gcccaccggc tacccagcct tcgaccacat acccaccggc
     1201 tccaactgcg cggcctgcgg ccttgcccca tcaatttttt taattttctc tggggaaaag
     1261 cctccggcct gcggcctgcg cgcttcgctt gccggttgga caccaagtgg aaggcgggtc
     1321 aaggctcgcg cagcgaccgc gcagcggctt ggccttgacg cgcctggaac gacccaagcc
     1381 tatgcgagtg ggggcagtcg aagggcgaag cccgcccgcc tgccccccga gcctcacggc
     1441 ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg caaggccgaa ggccgcgcag
     1501 tcgatcaaca agccccggag gggccacttt ttgccggagg gggagccgcg ccgaaggcgt
     1561 gggggaaccc cgcaggggtg cccttctttg ggcaccaaag aactagatat agggcgaaat
     1621 gcgaaagact taaaaatcaa caacttaaaa aaggggggta cgcaacagct cattgcggca
     1681 ccccccgcaa tagctcattg cgtaggttaa agaaaatctg taattgactg ccacttttac
     1741 gcaacgcata attgttgtcg cgctgccgaa aagttgcagc tgattgcgca tggtgccgca
     1801 accgtgcggc acccctaccg catggagata agcatggcca cgcagtccag agaaatcggc
     1861 attcaagcca agaacaagcc cggtcactgg gtgcaaacgg aacgcaaagc gcatgaggcg
     1921 tgggccgggc ttattgcgag gaaacccacg gcggcaatgc tgctgcatca cctcgtggcg
     1981 cagatgggcc accagaacgc cgtggtggtc agccagaaaa cactttccaa gctcatcgga
     2041 cgttctttgc ggacggtcca atacgcagtc aaggacttgg tggccgagcg ctggatctcc
     2101 gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg tggtcaatga ccgcgtggcg
     2161 tggggccagc cccgcgacca gttgcgcctg tcggtgttca gtgccgccgt ggtggttgat
     2221 cacgacgacc aggacgaatc gctgttgggg catggcgacc tgcgccgcat cccgaccctg
     2281 tatccgggcg agcagcaact accgaccggc cccggcgagg agccgcccag ccagcccggc
     2341 attccgggca tggaaccaga cctgccagcc ttgaccgaaa cggaggaatg ggaacggcgc
     2401 gggcagcagc gcctgccgat gcccgatgag ccgtgttttc tggacgatgg cgagccgttg
     2461 gagccgccga cacgggtaac gctgccgcgc cggtagggcc ggcctacggc cagcctcgca
     2521 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     2581 cgctcgcggg tgggcctact tcacctatcc tgcccttgtg tctcaaaatc tctgatgtac
     2641 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2701 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctccag gccgcgatta
     2761 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2821 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2881 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2941 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3001 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     3061 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3121 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3181 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3241 gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact
     3301 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3361 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3421 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3481 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta aggtaactgt
     3541 cagaccaagt ttactcatat atactttaga ttgatttttc gttccactga gcgtcagacc
     3601 ccagaactct gtcagaccga gtggaagaca t
//
