LOCUS       V043996                 8756 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043996
VERSION     V043996
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"
     regulatory      884..893
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             893..916
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             923..4243
                     /label="POLD1(NM_001256849)"
                     /note="POLD1(NM_001256849)"
                     /gene="POLD1"
     polyA_signal    4323..4547
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4593..5021
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5035..5364
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5431..6225
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6399..6520
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6569..6585)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6593..6609
                     /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(6617..6647)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6662..6683
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6971..7556)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7727..8587)
                     /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(8588..8692)
                     /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 ggctagcgtt taaacttaag cttgccacca tggactacaa
      901 ggacgacgat gacaagggat ccatggatgg caagcggcgg ccaggcccag ggcccggggt
      961 gcccccaaag cgggcccgtg ggggcctctg ggatgatgat gatgcacctc ggccatccca
     1021 attcgaggag gacctggcac tgatggagga gatggaggca gaacacaggc tgcaggagca
     1081 ggaggaggag gagctgcagt cagtcctgga gggggttgca gacgggcagg tcccaccatc
     1141 agccatagat cctcgctggc ttcggcccac accaccagcg ctggaccccc agacagagcc
     1201 cctcatcttc caacagttgg agattgacca ttatgtgggc ccagcgcagc ctgtgcctgg
     1261 ggggccccca ccatcccgcg gctccgtgcc tgtgctccgc gccttcgggg tcaccgatga
     1321 ggggttctct gtctgctgcc acatccacgg cttcgctccc tacttctaca ccccagcgcc
     1381 ccctggtttc gggcccgagc acatgggtga cctgcaacgg gagctgaact tggccatcag
     1441 ccgggacagt cgcgggggga gggagctgac tgggccggcc gtgctggctg tggaactgtg
     1501 ctcccgagag agcatgtttg ggtaccacgg gcacggcccc tccccgttcc tgcgcatcac
     1561 cgtggcgctg ccgcgcctcg tggccccggc ccgccgtctc ctggaacagg gcatccgtgt
     1621 ggcaggcctg ggcacgccca gcttcgcgcc ctacgaggcc aacgtcgact ttgagatccg
     1681 gttcatggtg gacacggaca tcgtcggctg caactggctg gagctcccag ctgggaaata
     1741 cgccctgagg ctgaaggaga aggctacgca gtgccagctg gaggcggacg tgctgtggtc
     1801 tgacgtggtc agtcacccac cggaagggcc atggcagcgc attgcgccct tgcgcgtgct
     1861 cagcttcgat atcgagtgcg ccggccgcaa aggcatcttc cctgagcctg agcgggaccc
     1921 tgtcatccag atctgctcgc tgggcctgcg ctggggggag ccggagccct tcctacgcct
     1981 ggcgctcacc ctgcggccct gtgcccccat cctgggtgcc aaggtgcaga gctacgagaa
     2041 ggaggaggac ctgctgcagg cctggtccac cttcatccgt atcatggacc ccgacgtgat
     2101 caccggttac aacatccaga acttcgacct tccgtacctc atctctcggg cccagaccct
     2161 caaggtacaa acattccctt tcctgggccg tgtggccggc ctttgctcca acatccggga
     2221 ctcttcattc cagtccaagc agacgggccg gcgggacacc aaggttgtca gcatggtggg
     2281 ccgcgtgcag atggacatgc tgcaggtgct gctgcgggag tacaagctcc gctcctacac
     2341 gctcaatgcc gtgagcttcc acttcctggg cgagcagaag gaggacgtgc agcacagcat
     2401 catcaccgac ctgcagaatg ggaacgacca gacccgccgc cgcctggctg tgtactgcct
     2461 gaaggatgcc tacctgccac tgcggctgct ggagcggctc atggtgctgg tgaacgccgt
     2521 ggagatggcg agggtcactg gcgtgcccct cagctacctg ctcagtcgtg gccagcaggt
     2581 caaggtcgta tcccagctgt tgcggcaggc catgcacgag gggctgctga tgcccgtggt
     2641 gaagtcagag ggcggcgagg actacacggg agccactgtc atcgagcccc tcaaagggta
     2701 ctacgacgtc cccatcgcca ccctggactt ctcctcgctg tacccgtcca tcatgatggc
     2761 ccacaacctg tgttacacca cgctccttcg gcccgggact gcacagaaac tgggcctgac
     2821 tgaggatcag ttcatcagga cccccaccgg ggacgagttt gtgaagacct cagtgcggaa
     2881 ggggctgctg ccccagatcc tggagaacct gctcagtgcc cggaagaggg ccaaggccga
     2941 gctggccaag gagacagacc ccctccggcg ccaggtcctg gatggacggc agctggcgct
     3001 gaaggtgagc gccaactccg tatacggctt cactggcgcc caggtgggca agttgccgtg
     3061 cctggagatc tcacagagcg tcacggggtt cggacgtcag atgatcgaga aaaccaagca
     3121 gctggtggag tctaagtaca cagtggagaa tggctacagc accagtgcca aggtggtgta
     3181 tggtgacact gactccgtca tgtgccgatt cggcgtgtcc tcggtggctg aggcgatggc
     3241 cctggggcgg gaggccgcgg actgggtgtc aggtcacttc ccgtcgccca tccggctgga
     3301 gtttgagaag gtctacttcc catacctgct tatcagcaag aagcgctacg cgggcctgct
     3361 cttctcctcc cggcccgacg cccacgaccg catggactgc aagggcctgg aggccgtgcg
     3421 cagggacaac tgccccctcg tggccaacct ggtcactgcc tcactgcgcc gcctgctcat
     3481 cgaccgagac cctgagggcg cggtggctca cgcacaggac gtcatctcgg acctgctgtg
     3541 caaccgcatc gatatctccc agctggtcat caccaaggag ctgacccgcg cggcctccga
     3601 ctatgccggc aagcaggccc acgtggagct ggccgagagg atgaggaagc gggaccccgg
     3661 gagtgcgccc agcctgggcg accgcgtccc ctacgtgatc atcagtgccg ccaagggtgt
     3721 ggccgcctac atgaagtcgg aggacccgct gttcgtgctg gagcacagcc tgcccattga
     3781 cacgcagtac tacctggagc agcagctggc caagcccctc ctgcgcatct tcgagcccat
     3841 cctgggcgag ggccgtgccg aggctgtgct actgcggggg gaccacacgc gctgcaagac
     3901 ggtgctcacg ggcaaggtgg gcggcctcct ggccttcgcc aaacgccgca actgctgcat
     3961 tggctgccgc acagtgctca gccaccaggg agccgtgtgt gagttctgcc agccccggga
     4021 gtctgagctg tatcagaagg aggtatccca tctgaatgcc ctggaggagc gcttctcgcg
     4081 cctctggacg cagtgccagc gctgccaggg cagcctgcac gaggacgtca tctgcaccag
     4141 ccgggactgc cccatcttct acatgcgcaa gaaggtgcgg aaggacctgg aagaccagga
     4201 gcagctcctg cggcgcttcg gaccccctgg acctgaggcc tggtgagaat tctgcagata
     4261 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     4321 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4381 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4441 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4501 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4561 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4801 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4921 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     5041 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5161 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5221 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5281 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5341 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5401 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5461 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5521 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5581 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5641 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5701 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5761 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5821 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5881 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5941 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6001 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6061 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6121 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6181 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6241 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6301 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6361 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6541 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6601 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6661 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6721 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6781 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6841 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6901 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7501 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8701 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
