LOCUS       V029476                 7382 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029476
VERSION     V029476
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..1579
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1616..3013
                     /label="VIM(NM_003380)"
                     /note="VIM(NM_003380)"
                     /gene="VIM"
     CDS             3020..3046
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3050..3076
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3080..3106
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3144..3368
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3414..3842
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3856..4185
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4252..4851
                     /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    5025..5146
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5195..5211)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5219..5235
                     /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(5243..5273)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5288..5309
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5597..6182)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6353..7213)
                     /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(7214..7318)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catggtgagc aagggcgagg aggataacat
      901 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      961 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1021 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1081 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1141 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1201 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1261 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1321 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1381 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1441 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1501 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1561 catggacgag ctgtacaagg gaggtggagg tagtggagga ggcggatcaa gcttcatgtc
     1621 caccaggtcc gtgtcctcgt cctcctaccg caggatgttc ggcggcccgg gcaccgcgag
     1681 ccggccgagc tccagccgga gctacgtgac tacgtccacc cgcacctaca gcctgggcag
     1741 cgcgctgcgc cccagcacca gccgcagcct ctacgcctcg tccccgggcg gcgtgtatgc
     1801 cacgcgctcc tctgccgtgc gcctgcggag cagcgtgccc ggggtgcggc tcctgcagga
     1861 ctcggtggac ttctcgctgg ccgacgccat caacaccgag ttcaagaaca cccgcaccaa
     1921 cgagaaggtg gagctgcagg agctgaatga ccgcttcgcc aactacatcg acaaggtgcg
     1981 cttcctggag cagcagaata agatcctgct ggccgagctc gagcagctca agggccaagg
     2041 caagtcgcgc ctgggggacc tctacgagga ggagatgcgg gagctgcgcc ggcaggtgga
     2101 ccagctaacc aacgacaaag cccgcgtcga ggtggagcgc gacaacctgg ccgaggacat
     2161 catgcgcctc cgggagaaat tgcaggagga gatgcttcag agagaggaag ccgaaaacac
     2221 cctgcaatct ttcagacagg atgttgacaa tgcgtctctg gcacgtcttg accttgaacg
     2281 caaagtggaa tctttgcaag aagagattgc ctttttgaag aaactccacg aagaggaaat
     2341 ccaggagctg caggctcaga ttcaggaaca gcatgtccaa atcgatgtgg atgtttccaa
     2401 gcctgacctc acggctgccc tgcgtgacgt acgtcagcaa tatgaaagtg tggctgccaa
     2461 gaacctgcag gaggcagaag aatggtacaa atccaagttt gctgacctct ctgaggctgc
     2521 caaccggaac aatgacgccc tgcgccaggc aaagcaggag tccactgagt accggagaca
     2581 ggtgcagtcc ctcacctgtg aagtggatgc ccttaaagga accaatgagt ccctggaacg
     2641 ccagatgcgt gaaatggaag agaactttgc cgttgaagct gctaactacc aagacactat
     2701 tggccgcctg caggatgaga ttcagaatat gaaggaggaa atggctcgtc accttcgtga
     2761 ataccaagac ctgctcaatg ttaagatggc ccttgacatt gagattgcca cctacaggaa
     2821 gctgctggaa ggcgaggaga gcaggatttc tctgcctctt ccaaactttt cctccctgaa
     2881 cctgagggaa actaatctgg attcactccc tctggttgat acccactcaa aaaggacact
     2941 tctgattaag acggttgaaa ctagagatgg acaggttatc aacgaaactt ctcagcatca
     3001 cgatgacctt gaactcgagt acccatacga cgtacctgat tatgcaactt atccttatga
     3061 tgtacctgat tatgctagct atccttatga tgtcccagac tacgcatagt gagggcccgt
     3121 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3181 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3241 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3301 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3361 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3541 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3781 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3841 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4021 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4081 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4141 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4201 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgaccgag
     4261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     4321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     4381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4621 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agcgggactc
     4861 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4921 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4981 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5161 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5221 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5281 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5341 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5401 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5461 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5521 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5581 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5641 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5701 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5761 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5821 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5881 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5941 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6001 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6061 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6121 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7321 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7381 gt
//
