LOCUS       V020743                 6716 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020743
VERSION     V020743
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             908..2209
                     /label="ACAT1(NM_001386677)"
                     /note="ACAT1(NM_001386677)"
                     /gene="ACAT1"
     CDS             2216..2242
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2283..2507
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2553..2981
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2995..3324
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3391..4185
                     /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    4359..4480
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4529..4545)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4553..4569
                     /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(4577..4607)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4622..4643
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4931..5516)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5687..6547)
                     /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(6548..6652)
                     /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 cttggtaccg agctcggatc
      901 cgccaccatg gctgtgctgg cggcacttct gcgcagcggc gcccgcagcc gcagccccct
      961 gctccggagg ctggtgcagg aaataagata tgtggaacgg agttatgtat caaaacccac
     1021 tttgaaggaa gtggtcatag taagtgctac aagaacaccc attggatctt ttttaggcag
     1081 cctttccttg ctgccagcca ctaagcttgg ttccattgca attcagggag ccattgaaaa
     1141 ggcagggatt ccaaaagaag aagtgaaaga agcatacatg ggtaatgttc tacaaggagg
     1201 tgaaggacaa gctcctacaa ggcaggcagt attgggtgca ggcttaccta tttctactcc
     1261 atgtaccacc ataaacaaag tttgtgcttc aggaatgaaa gccatcatga tggcctctca
     1321 aagtcttatg tgtggacatc aggatgtgat ggtggcaggt gggatggaga gcatgtccaa
     1381 tgttccatat gtaatgaaca gaggatcaac accatatggt ggggtaaagc ttgaagattt
     1441 gattgtaaaa gacgggctaa ctgatgtcta caataaaatt catatgggca gctgtgctga
     1501 gaatacagca aagaagctga atattgcacg aaatgaacag gacgcttatg ctattaattc
     1561 ttataccaga agtaaagcag catgggaagc tgggaaattt ggaaatgaag ttattcctgt
     1621 cacagttaca gtaaaaggtc aaccagatgt agtggtgaaa gaagatgaag aatataaacg
     1681 tgttgatttt agcaaagttc caaagctgaa gacagttttc cagaaagaaa atggcacagt
     1741 aacagctgcc aatgccagta cactgaatga tggagcagct gctctggttc tcatgacggc
     1801 agatgcagcg aagaggctca atgttacacc actggcaaga atagtagcat ttgctgacgc
     1861 tgctgtagaa cctattgatt ttccaattgc tcctgtatat gctgcatcta tggttcttaa
     1921 agatgtggga ttgaaaaaag aagatattgc aatgtgggaa gtaaatgaag cctttagtct
     1981 ggttgtacta gcaaacatta aaatgttgga gattgatccc caaaaagtga atatcaatgg
     2041 aggagctgtt tctctgggac atccaattgg taaatgcttt cttaatttta ggatgtctgg
     2101 agccaggatt gttggtcatt tgactcatgc cttgaagcaa ggagaatacg gtcttgccag
     2161 tatttgcaat ggaggaggag gtgcttctgc catgctaatt cagaagctgc tcgagtaccc
     2221 atacgacgta ccagattacg cttagtctag agggcccgtt taaacccgct gatcagcctc
     2281 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2341 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2401 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2461 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2521 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2701 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2761 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2881 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3001 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3061 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3121 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3181 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3241 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3301 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3361 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3421 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3481 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3541 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3601 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3661 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3721 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3781 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3841 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3901 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3961 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4021 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4081 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4141 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4201 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4261 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4321 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4441 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4501 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4561 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4621 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4681 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4741 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4801 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4861 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5341 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5461 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5641 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5701 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5761 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5821 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5881 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5941 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6001 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6061 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6121 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6181 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6241 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6301 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6361 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6421 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6481 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6541 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6661 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
