LOCUS       sequence-388772-        3695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of J23100 and BCD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 5 to 39)
  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 3695)
  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(J23100-BCD2-E0040m-B0015)
FEATURES             Location/Qualifiers
     source          1..3695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             114..122
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             132..845
                     /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(162..190)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     785..805
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      864..935
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      951..978
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      1126..1897
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1898..2560
                     /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            2571..2679
                     /note="incP origin of transfer"
     primer_bind     complement(2769..2788)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2780..3595
                     /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             2780..3595
                     /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(2853..2872)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggagttgacg gctagctcag tcctaggtac agtgctagct actgggccca agttcactta
       61 aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca tgctaaggag
      121 gttttctaat gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      181 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      241 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
      301 ggccaacact tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc
      361 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
      421 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
      481 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
      541 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
      601 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
      661 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
      721 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagatc
      781 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
      841 acaaataata aaggtccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      901 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
      961 cgggtgggcc tttctgcgtt tatagcttat gtcttcagca gtaggacaaa tccgccgccc
     1021 tagacagctg ggcgcgcccc ccctacgggc ttgctctccg ggcttcgccc tgcgcggtcg
     1081 ctgcgctccc ttgccagccc gtggatatgt ggacgatggc cgcgagcggc caccggctgg
     1141 ctcgcttcgc tcggcccgtg gacaaccctg ctggacaagc tgatggacag gctgcgcctg
     1201 cccacgagct tgaccacagg gattgcccac cggctaccca gccttcgacc acatacccac
     1261 cggctccaac tgcgcggcct gcggccttgc cccatcaatt tttttaattt tctctgggga
     1321 aaagcctccg gcctgcggcc tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg
     1381 ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca
     1441 agcctatgcg agtgggggca gtcgaagggc gaagcccgcc cgcctgcccc ccgagcctca
     1501 cggcggcgag tgcgggggtt ccaagggggc agcgccacct tgggcaaggc cgaaggccgc
     1561 gcagtcgatc aacaagcccc ggaggggcca ctttttgccg gagggggagc cgcgccgaag
     1621 gcgtggggga accccgcagg ggtgcccttc tttgggcacc aaagaactag atatagggcg
     1681 aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac agctcattgc
     1741 ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg actgccactt
     1801 ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg cagctgattg cgcatggtgc
     1861 cgcaaccgtg cggcacccct accgcatgga gataagcatg gccacgcagt ccagagaaat
     1921 cggcattcaa gccaagaaca agcccggtca ctgggtgcaa acggaacgca aagcgcatga
     1981 ggcgtgggcc gggcttattg cgaggaaacc cacggcggca atgctgctgc atcacctcgt
     2041 ggcgcagatg ggccaccaga acgccgtggt ggtcagccag aaaacacttt ccaagctcat
     2101 cggacgttct ttgcggacgg tccaatacgc agtcaaggac ttggtggccg agcgctggat
     2161 ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc tacgtggtca atgaccgcgt
     2221 ggcgtggggc cagccccgcg accagttgcg cctgtcggtg ttcagtgccg ccgtggtggt
     2281 tgatcacgac gaccaggacg aatcgctgtt ggggcatggc gacctgcgcc gcatcccgac
     2341 cctgtatccg ggcgagcagc aactaccgac cggccccggc gaggagccgc ccagccagcc
     2401 cggcattccg ggcatggaac cagacctgcc agccttgacc gaaacggagg aatgggaacg
     2461 gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt tttctggacg atggcgagcc
     2521 gttggagccg ccgacacggg taacgctgcc gcgccggtag ggccggccta cggccagcct
     2581 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     2641 cacccgctcg cgggtgggcc tacttcacct atcctgccct tgtgtctcaa aatctctgat
     2701 gtacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2761 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg
     2821 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2881 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     2941 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3001 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3061 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     3121 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3181 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3241 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3301 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     3361 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3421 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3481 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3541 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaaggtaa
     3601 ctgtcagacc aagtttactc atatatactt tagattgatt tttcgttcca ctgagcgtca
     3661 gaccccagaa ctctgtcaga ccgagtggaa gacat
//
