LOCUS       sequence-37550-e        3848 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 3848)
  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 3848)
  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: pACBB-eGFP
FEATURES             Location/Qualifiers
     source          1..3848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(885..1544)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1459..1478
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1545..1647)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2039..2056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2173..2717)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2253..2272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2907..2928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2943..2948,2949..2966,2967..2973)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             join(3019..3021,3022..3024,3025..3738)
                     /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(3019..3021,3022..3024,3025..3735)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3064..3085)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3325..3344)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3672..3693
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3745..3754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3764..3781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
       61 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
      121 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
      181 ggtttccgtg tttcgtaaag tctggaaacg cggaagtccc ctacgtgctg ctgaagttgc
      241 ccgcaacaga gagtggaacc aaccggtgat accacgatac tatgactgag agtcaacgcc
      301 atgagcggcc tcatttctta ttctgagtta caacagtccg caccgctgtc cggtagctcc
      361 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
      421 caactcgtag gacaggtgcc ggcagcgccc aacagtcccc cggccacggg gcctgccacc
      481 atacccacgc cgaaacaagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
      541 tgctggctac cctgtggaac acctacatct gtattaacga agcgctaacc gtttttatca
      601 ggctctggga ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct
      661 gagcaaactg gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
      721 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
      781 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
      841 gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
      901 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacag
      961 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     1021 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     1081 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1141 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1201 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1261 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1321 atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     1381 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     1441 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     1501 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     1561 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     1621 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     1681 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     1741 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     1801 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     1861 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     1921 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     1981 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     2041 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     2101 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     2161 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     2221 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc ccctggcggc
     2281 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     2341 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     2401 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2521 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2581 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2641 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     2701 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc aagatttcag
     2761 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     2821 tctcatgttt gacagcttat catcgatgaa ttcatcctga acttatctag acccgactgg
     2881 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2941 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgt ctagtagaag
     3001 gaggagatct ggatccatat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3061 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3121 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3181 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3241 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3301 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3361 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3421 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3481 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3541 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3601 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3661 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3721 gacgagctgt acaagtaaat gcatccatgg cggccgcctc gagcatcacc atcaccatca
     3781 ctgattctcc ttacgcatct gtgcggtatt tcacaccgca tgtactagtt aacttcagga
     3841 cctgcagc
//
