LOCUS       V023055                 6843 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023055
VERSION     V023055
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"
     regulatory      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             943..2364
                     /label="HNF4A(NM_000457)"
                     /note="HNF4A(NM_000457)"
                     /gene="HNF4A"
     polyA_signal    2443..2667
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2713..3141
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3155..3484
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3551..4345
                     /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    4519..4640
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4689..4705)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4713..4729
                     /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(4737..4767)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4782..4803
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5091..5676)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5847..6707)
                     /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(6708..6812)
                     /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 gccaccatgg attacaagga tgacgacgat aagactaagc ttatgcgact ctccaaaacc
      961 ctcgtcgaca tggacatggc cgactacagt gctgcactgg acccagccta caccaccctg
     1021 gaatttgaga atgtgcaggt gttgacgatg ggcaatgaca cgtccccatc agaaggcacc
     1081 aacctcaacg cgcccaacag cctgggtgtc agcgccctgt gtgccatctg cggggaccgg
     1141 gccacgggca aacactacgg tgcctcgagc tgtgacggct gcaagggctt cttccggagg
     1201 agcgtgcgga agaaccacat gtactcctgc agatttagcc ggcagtgcgt ggtggacaaa
     1261 gacaagagga accagtgccg ctactgcagg ctcaagaaat gcttccgggc tggcatgaag
     1321 aaggaagccg tccagaatga gcgggaccgg atcagcactc gaaggtcaag ctatgaggac
     1381 agcagcctgc cctccatcaa tgcgctcctg caggcggagg tcctgtcccg acagatcacc
     1441 tcccccgtct ccgggatcaa cggcgacatt cgggcgaaga agattgccag catcgcagat
     1501 gtgtgtgagt ccatgaagga gcagctgctg gttctcgttg agtgggccaa gtacatccca
     1561 gctttctgcg agctccccct ggacgaccag gtggccctgc tcagagccca tgctggcgag
     1621 cacctgctgc tcggagccac caagagatcc atggtgttca aggacgtgct gctcctaggc
     1681 aatgactaca ttgtccctcg gcactgcccg gagctggcgg agatgagccg ggtgtccata
     1741 cgcatccttg acgagctggt gctgcccttc caggagctgc agatcgatga caatgagtat
     1801 gcctacctca aagccatcat cttctttgac ccagatgcca aggggctgag cgatccaggg
     1861 aagatcaagc ggctgcgttc ccaggtgcag gtgagcttgg aggactacat caacgaccgc
     1921 cagtatgact cgcgtggccg ctttggagag ctgctgctgc tgctgcccac cttgcagagc
     1981 atcacctggc agatgatcga gcagatccag ttcatcaagc tcttcggcat ggccaagatt
     2041 gacaacctgt tgcaggagat gctgctggga gggtccccca gcgatgcacc ccatgcccac
     2101 caccccctgc accctcacct gatgcaggaa catatgggaa ccaacgtcat cgttgccaac
     2161 acaatgccca ctcacctcag caacggacag atgtgtgagt ggccccgacc caggggacag
     2221 gcagccaccc ctgagacccc acagccctca ccgccaggtg gctcagggtc tgagccctat
     2281 aagctcctgc cgggagccgt cgccacaatc gtcaagcccc tctctgccat cccccagccg
     2341 accatcacca agcaggaagt tatctagaat tctgcagata tccagcacag tggcggccgc
     2401 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2461 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2521 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2581 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2641 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2701 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2761 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2821 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2881 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2941 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3001 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3061 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3121 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3301 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3361 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3421 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3481 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3541 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3601 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3661 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3721 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3781 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3841 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3901 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3961 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4021 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4081 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4141 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4201 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4261 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4321 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4381 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4441 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4501 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4621 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4681 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4741 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4921 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4981 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5041 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5101 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5161 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5221 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5281 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5341 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5401 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5461 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5521 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5581 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     5641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6841 gtc
//
