LOCUS       sequence-37553-e        4207 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 4207)
  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 4207)
  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: pUCBB-pBAD-eGFP
FEATURES             Location/Qualifiers
     source          1..4207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(85..102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(256..844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(336..355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1015..1806,1807..1875))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1638..1657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1876..1980)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2048..2066
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2070..2113)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(2140..3018)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3045..3329
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3252..3271
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(3380..3382,3383..3385,3386..4099)
                     /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(3380..3382,3383..3385,3386..4096)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3425..3446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3686..3705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4033..4054
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      4106..4115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4125..4142
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gtccggcaaa aaaacgggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
       61 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      661 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1981 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2041 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
     2101 taaggatgat ttctggaatt catcctgaac ttatctagat tatgacaact tgacggctac
     2161 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     2221 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     2281 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     2341 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     2401 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     2461 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     2521 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     2581 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     2641 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     2701 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     2761 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     2821 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     2881 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     2941 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     3001 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     3061 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     3121 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     3181 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     3241 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     3301 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa ataattttgt
     3361 ttaagagatc tggatccata tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3421 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3481 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3541 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3601 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3661 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3721 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3781 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3841 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3901 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3961 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4021 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4081 ggacgagctg tacaagtaaa tgcatccatg gcggccgcct cgagcatcac catcaccatc
     4141 actgattctc cttacgcatc tgtgcggtat ttcacaccgc atgtactagt taacttcagg
     4201 acctgca
//
