LOCUS       sequence-388744-        3633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of J23103 and B0032m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3633)
  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 3633)
  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-B0032m-E0040m-B0015)
FEATURES             Location/Qualifiers
     source          1..3633
                     /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             70..783
                     /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(100..128)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     723..743
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      802..873
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      889..916
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      1064..1835
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1836..2498
                     /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            2509..2617
                     /note="incP origin of transfer"
     primer_bind     complement(2707..2726)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2718..3533
                     /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             2718..3533
                     /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(2791..2810)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggagctgata gctagctcag tcctagggat tatgctagct actagagtca cacaggaaag
       61 tactaaatga tgcgtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      121 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      181 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      241 ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat
      301 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
      361 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
      421 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
      481 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
      541 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
      601 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
      661 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagatcac
      721 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
      781 aaataataaa ggtccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      841 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
      901 ggtgggcctt tctgcgttta tagcttatgt cttcagcagt aggacaaatc cgccgcccta
      961 gacagctggg cgcgcccccc ctacgggctt gctctccggg cttcgccctg cgcggtcgct
     1021 gcgctccctt gccagcccgt ggatatgtgg acgatggccg cgagcggcca ccggctggct
     1081 cgcttcgctc ggcccgtgga caaccctgct ggacaagctg atggacaggc tgcgcctgcc
     1141 cacgagcttg accacaggga ttgcccaccg gctacccagc cttcgaccac atacccaccg
     1201 gctccaactg cgcggcctgc ggccttgccc catcaatttt tttaattttc tctggggaaa
     1261 agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg
     1321 tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga acgacccaag
     1381 cctatgcgag tgggggcagt cgaagggcga agcccgcccg cctgcccccc gagcctcacg
     1441 gcggcgagtg cgggggttcc aagggggcag cgccaccttg ggcaaggccg aaggccgcgc
     1501 agtcgatcaa caagccccgg aggggccact ttttgccgga gggggagccg cgccgaaggc
     1561 gtgggggaac cccgcagggg tgcccttctt tgggcaccaa agaactagat atagggcgaa
     1621 atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg tacgcaacag ctcattgcgg
     1681 caccccccgc aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac tgccactttt
     1741 acgcaacgca taattgttgt cgcgctgccg aaaagttgca gctgattgcg catggtgccg
     1801 caaccgtgcg gcacccctac cgcatggaga taagcatggc cacgcagtcc agagaaatcg
     1861 gcattcaagc caagaacaag cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg
     1921 cgtgggccgg gcttattgcg aggaaaccca cggcggcaat gctgctgcat cacctcgtgg
     1981 cgcagatggg ccaccagaac gccgtggtgg tcagccagaa aacactttcc aagctcatcg
     2041 gacgttcttt gcggacggtc caatacgcag tcaaggactt ggtggccgag cgctggatct
     2101 ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta cgtggtcaat gaccgcgtgg
     2161 cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg
     2221 atcacgacga ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc
     2281 tgtatccggg cgagcagcaa ctaccgaccg gccccggcga ggagccgccc agccagcccg
     2341 gcattccggg catggaacca gacctgccag ccttgaccga aacggaggaa tgggaacggc
     2401 gcgggcagca gcgcctgccg atgcccgatg agccgtgttt tctggacgat ggcgagccgt
     2461 tggagccgcc gacacgggta acgctgccgc gccggtaggg ccggcctacg gccagcctcg
     2521 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     2581 cccgctcgcg ggtgggccta cttcacctat cctgcccttg tgtctcaaaa tctctgatgt
     2641 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     2701 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcc aggccgcgat
     2761 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2821 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     2881 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2941 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     3001 ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg
     3061 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     3121 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     3181 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     3241 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     3301 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     3361 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     3421 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     3481 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taaggtaact
     3541 gtcagaccaa gtttactcat atatacttta gattgatttt tcgttccact gagcgtcaga
     3601 ccccagaact ctgtcagacc gagtggaaga cat
//
