LOCUS       sequence-37548-e        4857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4857)
  AUTHORS   Vick JE, Johnson ET, Choudhary S, Bloch SE, Lopez-Gallego F, 
            Srivastava P, Tikh IB, Wawrzyn GT, Schmidt-Dannert C
  TITLE     Optimized compatible set of BioBrick vectors for metabolic pathway 
            engineering.
  JOURNAL   Appl Microbiol Biotechnol. 2011 Dec;92(6):1275-86. Epub 2011 Oct 28.
  PUBMED    22033566
REFERENCE   2  (bases 1 to 4857)
  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: "Appl 
            Microbiol Biotechnol."; date: "2011-12"; volume: "92(6)"; pages: 
            "1275-86. Epub 2011 Oct 28"
COMMENT     Sequence Label: pBBRBB-eGFP
FEATURES             Location/Qualifiers
     source          1..4857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(51..713)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      714..1483
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2963..3757
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3017..3036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3627..3646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    3916..3937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3952..3957,3958..3975,3976..3982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             join(4028..4030,4031..4033,4034..4747)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4028..4030,4031..4033,4034..4744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4073..4094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4334..4353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4681..4702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      4754..4763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4773..4790
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctagtatagt ctggaacagc gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc
       61 ggcagcgtga cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca
      121 tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct
      181 ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg
      241 gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc
      301 aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg
      361 cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg
      421 gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact
      481 gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc
      541 accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg
      601 ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag
      661 tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc
      721 tccatgcggt agggtgccgc acggttgcgg caccatgcgc aatcagctgc aacttttcgg
      781 cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc aattacagat tttctttaac
      841 ctacgcaatg agctattgcg gggggtgccg caatgagctg ttgcgtaccc ccctttttta
      901 agttgttgat ttttaagtct ttcgcatttc gccctatatc tagttctttg gtgcccaaag
      961 aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg ctccccctcc ggcaaaaagt
     1021 ggcccctccg gggcttgttg atcgactgcg cggccttcgg ccttgcccaa ggtggcgctg
     1081 cccccttgga acccccgcac tcgccgccgt gaggctcggg gggcaggcgg gcgggcttcg
     1141 ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg
     1201 ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga
     1261 agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa ttgatggggc
     1321 aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg ggtagccggt
     1381 gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca
     1441 gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc catcgtccac
     1501 atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg gagagcaagc
     1561 ccgtagggcg ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag tctagtgagt
     1621 atactcaagc attgagtggc ccgccggagg caccgccttg cgctgccccc gtcgagccgg
     1681 ttggacacca aaagggaggg gcaggcatgg cggcatacgc gatcatgcga tgcaagaagc
     1741 tggcgaaaat gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag cgcgagacgc
     1801 ccaacgctga cgccagcagg acgccagaga acgagcactg ggcggccagc agcaccgatg
     1861 aagcgatggg ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac gctgtgttgg
     1921 cggtcgagta cgtcatgacg gccagcccgg aatggtggaa gtcggccagc caagaacagc
     1981 aggcggcgtt cttcgagaag gcgcacaagt ggctggcgga caagtacggg gcggatcgca
     2041 tcgtgacggc cagcatccac cgtgacgaaa ccagcccgca catgaccgcg ttcgtggtgc
     2101 cgctgacgca ggacggcagg ctgtcggcca aggagttcat cggcaacaaa gcgcagatga
     2161 cccgcgacca gaccacgttt gcggctgctg tggccgatct agggctgcaa cggggcatcg
     2221 agggcagcaa ggcacgtcac acgcgcattc aggcgttcta cgaggccctg gagcggccac
     2281 cagtgggcca cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat gcaccgcagg
     2341 gattggccga aaagctggga atctcaaagc gcgttgagac gccggaagcc gtggccgacc
     2401 ggctgacaaa agcggttcgg caggggtatg agcctgccct acaggccgcc gcaggagcgc
     2461 gtgagatgcg caagaaggcc gatcaagccc aagagacggc ccgagacctt cgggagcgcc
     2521 tgaagcccgt tctggacgcc ctggggccgt tgaatcggga tatgcaggcc aaggccgccg
     2581 cgatcatcaa ggccgtgggc gaaaagctgc tgacggaaca gcgggaagtc cagcgccaga
     2641 aacaggccca gcgccagcag gaacgcgggc gcgcacattt ccccgaaaag tgccacctgg
     2701 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
     2761 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     2821 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     2881 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ttgatcaaga
     2941 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3001 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3061 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3121 gtccggtgcc ctgaatgaac tacaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3181 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3241 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3301 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3361 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3421 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3481 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3541 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3601 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3661 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3721 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3781 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3841 gaaaggttgg actagagaat tcatcctgaa cttatctaga cccgactgga aagcgggcag
     3901 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3961 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgtc tagtagaagg aggagatctg
     4021 gatccatatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4081 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4141 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     4201 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     4261 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     4321 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     4381 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     4441 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     4501 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     4561 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4621 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     4681 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4741 caagtaaatg catccatggc ggccgcctcg agcatcacca tcaccatcac tgattctcct
     4801 tacgcatctg tgcggtattt cacaccgcat gtactagtta acttcaggac ctgcagt
//
