LOCUS       V038708                 6595 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038708
VERSION     V038708
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             937..1653
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1690..2346
                     /label="STX4(NM_001272095)"
                     /note="STX4(NM_001272095)"
                     /gene="STX4"
     CDS             2359..2424
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2459..2683
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2729..3157
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3171..3500
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3512..4111
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4271..4392
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4441..4457)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4465..4481
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4489..4519)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4534..4555
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4843..5428)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5599..6459)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6460..6564)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcga tatcggatcc gccaccatgg tgagcaaggg cgaggagctg
      961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1621 gggatcactc tcggcatgga cgagctgtac aagggcggtg gaggcagtgg tggaggcggt
     1681 agcggtacca tgaagcagga gctgcagaac ctgcgcgatg agatcaaaca gctggggagg
     1741 gagatccgcc tgcagctgaa ggccatagag ccccagaagg aggaagctga tgagaactat
     1801 aactccgtca acacaagaat gagaaaaacc cagcatgggg tcctgtccca gcaattcgtg
     1861 gagctcatca acaagtgcaa ttcaatgcag tccgaatacc gggagaagaa cgtggagcgg
     1921 attcggaggc agctgaagat caccaatgct gggatggtgt ctgatgagga gttggagcag
     1981 atgctggaca gtgggcaaag cgaggtgttt gtgtccaata tcctgaagga cacgcaggtg
     2041 actcgacagg ccttaaatga gatctcggcc cggcacagtg agatccagca gcttgaacgc
     2101 agtattcgtg agctgcacga catattcact tttctggcta ccgaagtgga gatgcagggg
     2161 gagatgatca atcggattga gaagaacatc ctgagctcag cggactacgt ggaacgtggg
     2221 caggagcacg tcaagacggc cctggagaac cagaagaagg cgaggaagaa gaaagtcttg
     2281 attgccatct gtgtgtccat caccgtcgtc ctcctagcag tcatcattgg cgtcacagtg
     2341 gttggactcg agaccggtga ctacaaagac catgacggtg attataaaga tcatgacatc
     2401 gattacaagg atgacgatga caagtgaccg gtaagtttaa accgctgatc agcctcgact
     2461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2641 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2701 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3061 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta
     3121 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3421 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3481 gaggcctagg cttttgcaaa aagctcccgg gatgaccgag tacaagccca cggtgcgcct
     3541 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     3601 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     3661 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     3721 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     3781 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     3841 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     3901 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     3961 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4021 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4081 ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa
     4141 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4201 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4261 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4321 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4381 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
