LOCUS       sequence-37549-e        2939 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 2939)
  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 2939)
  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: pCDFBB-eGFP
FEATURES             Location/Qualifiers
     source          1..2939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..760
                     /direction=RIGHT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     promoter        808..899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             900..1691
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(1862..1909)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1905..1923
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    1998..2019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2034..2039,2040..2057,2058..2064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             join(2110..2112,2113..2115,2116..2829)
                     /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(2110..2112,2113..2115,2116..2826)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2155..2176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2416..2435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2763..2784
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2836..2845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2855..2872
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 cttgagcggt tcagtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
       61 aatcttttgc cctgtaaacg aaaaaaccac ctggggaggt ggtttgatcg aaggttaagt
      121 cagttgggga actgcttaac cgtggtaact ggctttcgca gagcacagca accaaatctg
      181 tccttccagt gtagccggac tttggcgcac acttcaagag caaccgcgtg tttagctaaa
      241 caaatcctct gcgaactccc agttaccaat ggctgctgcc agtggcgttt taccgtgctt
      301 ttccgggttg gactcaagtg aacagttacc ggataaggcg cagcagtcgg gctgaacggg
      361 gagttcttgc ttacagccca gcttggagcg aacgacctac accgagccga gataccagtg
      421 tgtgagctat gagaaagcgc cacacttccc gtaagggaga aaggcggaac aggtatccgg
      481 taaacggcag ggtcggaaca ggagagcgca agagggagcg acccgccgga aacggtgggg
      541 atctttaagt cctgtcgggt ttcgcccgta ctgtcagatt catggttgag cctcacggct
      601 cccacagatg caccggaaaa gcgtctgttt atgtgaactc tggcaggagg gcggagccta
      661 tggaaaaacg ccaccggcgc ggccctgctg ttttgcctca catgttagtc ccctgcttat
      721 ccacggaatc tgtgggtaac tttgtatgtg tccgcagcgc ccgccgcagt ctcacgcccg
      781 gagcgtagcg accgagtgag ctagctattt gtttattttt ctaaatacat tcaaatatgt
      841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      901 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
      961 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     1021 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     1081 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     1141 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     1201 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     1261 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     1321 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     1381 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     1441 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     1501 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     1561 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     1621 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     1681 tcggcaaata atgtctaaca attcgttcaa gccgaggggc cgcaagatcc ggccacgatg
     1741 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
     1801 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggtttca
     1861 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
     1921 cggtggcagc agcctaggga attcatcctg aacttatcta gacccgactg gaaagcgggc
     1981 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     2041 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg tctagtagaa ggaggagatc
     2101 tggatccata tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2161 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2221 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2281 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2341 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2401 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2461 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2521 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2581 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2641 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2701 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2761 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2821 tacaagtaaa tgcatccatg gcggccgcct cgagcatcac catcaccatc actgattctc
     2881 cttacgcatc tgtgcggtat ttcacaccgc atgtactagt taacttcagg acctgcagt
//
