LOCUS       V038333                 9145 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038333
VERSION     V038333
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..4896
                     /label="TERT(NM_001193376)"
                     /note="TERT(NM_001193376)"
                     /gene="TERT"
     CDS             4909..4974
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5009..5233
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5279..5707
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5721..6050
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6062..6661
                     /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    6821..6942
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6991..7007)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7015..7031
                     /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(7039..7069)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7084..7105
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7393..7978)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8149..9009)
                     /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(9010..9114)
                     /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 tgccgcgcgc tccccgctgc cgagccgtgc gctccctgct gcgcagccac
     1741 taccgcgagg tgctgccgct ggccacgttc gtgcggcgcc tggggcccca gggctggcgg
     1801 ctggtgcagc gcggggaccc ggcggctttc cgcgcgctgg tggcccagtg cctggtgtgc
     1861 gtgccctggg acgcacggcc gccccccgcc gccccctcct tccgccaggt gtcctgcctg
     1921 aaggagctgg tggcccgagt gctgcagagg ctgtgcgagc gcggcgcgaa gaacgtgctg
     1981 gccttcggct tcgcgctgct ggacggggcc cgcgggggcc cccccgaggc cttcaccacc
     2041 agcgtgcgca gctacctgcc caacacggtg accgacgcac tgcgggggag cggggcgtgg
     2101 gggctgctgc tgcgccgcgt gggcgacgac gtgctggttc acctgctggc acgctgcgcg
     2161 ctctttgtgc tggtggctcc cagctgcgcc taccaggtgt gcgggccgcc gctgtaccag
     2221 ctcggcgctg ccactcaggc ccggcccccg ccacacgcta gtggaccccg aaggcgtctg
     2281 ggatgcgaac gggcctggaa ccatagcgtc agggaggccg gggtccccct gggcctgcca
     2341 gccccgggtg cgaggaggcg cgggggcagt gccagccgaa gtctgccgtt gcccaagagg
     2401 cccaggcgtg gcgctgcccc tgagccggag cggacgcccg ttgggcaggg gtcctgggcc
     2461 cacccgggca ggacgcgtgg accgagtgac cgtggtttct gtgtggtgtc acctgccaga
     2521 cccgccgaag aagccacctc tttggagggt gcgctctctg gcacgcgcca ctcccaccca
     2581 tccgtgggcc gccagcacca cgcgggcccc ccatccacat cgcggccacc acgtccctgg
     2641 gacacgcctt gtcccccggt gtacgccgag accaagcact tcctctactc ctcaggcgac
     2701 aaggagcagc tgcggccctc cttcctactc agctctctga ggcccagcct gactggcgct
     2761 cggaggctcg tggagaccat ctttctgggt tccaggccct ggatgccagg gactccccgc
     2821 aggttgcccc gcctgcccca gcgctactgg caaatgcggc ccctgtttct ggagctgctt
     2881 gggaaccacg cgcagtgccc ctacggggtg ctcctcaaga cgcactgccc gctgcgagct
     2941 gcggtcaccc cagcagccgg tgtctgtgcc cgggagaagc cccagggctc tgtggcggcc
     3001 cccgaggagg aggacacaga cccccgtcgc ctggtgcagc tgctccgcca gcacagcagc
     3061 ccctggcagg tgtacggctt cgtgcgggcc tgcctgcgcc ggctggtgcc cccaggcctc
     3121 tggggctcca ggcacaacga acgccgcttc ctcaggaaca ccaagaagtt catctccctg
     3181 gggaagcatg ccaagctctc gctgcaggag ctgacgtgga agatgagcgt gcgggactgc
     3241 gcttggctgc gcaggagccc aggggttggc tgtgttccgg ccgcagagca ccgtctgcgt
     3301 gaggagatcc tggccaagtt cctgcactgg ctgatgagtg tgtacgtcgt cgagctgctc
     3361 aggtctttct tttatgtcac ggagaccacg tttcaaaaga acaggctctt tttctaccgg
     3421 aagagtgtct ggagcaagtt gcaaagcatt ggaatcagac agcacttgaa gagggtgcag
     3481 ctgcgggagc tgtcggaagc agaggtcagg cagcatcggg aagccaggcc cgccctgctg
     3541 acgtccagac tccgcttcat ccccaagcct gacgggctgc ggccgattgt gaacatggac
     3601 tacgtcgtgg gagccagaac gttccgcaga gaaaagaggg ccgagcgtct cacctcgagg
     3661 gtgaaggcac tgttcagcgt gctcaactac gagcgggcgc ggcgccccgg cctcctgggc
     3721 gcctctgtgc tgggcctgga cgatatccac agggcctggc gcaccttcgt gctgcgtgtg
     3781 cgggcccagg acccgccgcc tgagctgtac tttgtcaagg tggatgtgac gggcgcgtac
     3841 gacaccatcc cccaggacag gctcacggag gtcatcgcca gcatcatcaa accccagaac
     3901 acgtactgcg tgcgtcggta tgccgtggtc cagaaggccg cccatgggca cgtccgcaag
     3961 gccttcaaga gccacgtctc taccttgaca gacctccagc cgtacatgcg acagttcgtg
     4021 gctcacctgc aggagaccag cccgctgagg gatgccgtcg tcatcgagca gagctcctcc
     4081 ctgaatgagg ccagcagtgg cctcttcgac gtcttcctac gcttcatgtg ccaccacgcc
     4141 gtgcgcatca ggggcaagtc ctacgtccag tgccagggga tcccgcaggg ctccatcctc
     4201 tccacgctgc tctgcagcct gtgctacggc gacatggaga acaagctgtt tgcggggatt
     4261 cggcgggacg ggctgctcct gcgtttggtg gatgatttct tgttggtgac acctcacctc
     4321 acccacgcga aaaccttcct cagctatgcc cggacctcca tcagagccag tctcaccttc
     4381 aaccgcggct tcaaggctgg gaggaacatg cgtcgcaaac tctttggggt cttgcggctg
     4441 aagtgtcaca gcctgtttct ggatttgcag gtgaacagcc tccagacggt gtgcaccaac
     4501 atctacaaga tcctcctgct gcaggcgtac aggtttcacg catgtgtgct gcagctccca
     4561 tttcatcagc aagtttggaa gaaccccaca tttttcctgc gcgtcatctc tgacacggcc
     4621 tccctctgct actccatcct gaaagccaag aacgcaggga tgtcgctggg ggccaagggc
     4681 gccgccggcc ctctgccctc cgaggccgtg cagtggctgt gccaccaagc attcctgctc
     4741 aagctgactc gacaccgtgt cacctacgtg ccactcctgg ggtcactcag gacagcccag
     4801 acgcagctga gtcggaagct cccggggacg acgctgactg ccctggaggc cgcagccaac
     4861 ccggcactgc cctcagactt caagaccatc ctggacctcg agaccggtga ctacaaagac
     4921 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagtgaccg
     4981 gtaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     5041 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     5101 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5161 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5221 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     5281 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5341 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5401 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5461 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5521 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5581 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5641 gggattttgg ccatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5701 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     6001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     6061 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     6121 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     6181 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     6241 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     6301 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     6361 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     6421 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     6481 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     6541 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     6601 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6661 attcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6721 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6781 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6841 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6901 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6961 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7021 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7081 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7141 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7201 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7261 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7321 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7381 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7441 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7501 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7561 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7621 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7681 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7741 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7801 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7861 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7921 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9121 tccccgaaaa gtgccacctg acgtc
//
