LOCUS       V019577                 7031 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019577
VERSION     V019577
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..1690
                     /label="RNF2(NM_007212)"
                     /note="RNF2(NM_007212)"
                     /gene="RNF2"
     CDS             1697..1723
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1727..1753
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1757..1783
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    1807..2395
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2461..2509
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2711..3139
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3153..3482
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3549..4343
                     /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..5679)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5850..6710)
                     /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(6711..6815)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgtctca ggctgtgcag acaaacggaa ctcaaccatt
      721 aagcaaaaca tgggaactca gtttatatga gttacaacga acacctcagg aggcaataac
      781 agatggctta gaaattgtgg tttcacctcg aagtctacac agtgaattaa tgtgcccaat
      841 ttgtttggat atgttgaaga acaccatgac tacaaaggag tgtttacatc gtttttgtgc
      901 agactgcatc atcacagccc ttagaagtgg caacaaagaa tgtcctacct gtcggaaaaa
      961 actagtttcc aaaagatcac taaggccaga cccaaacttt gatgcactca tcagcaaaat
     1021 ttatccaagt cgtgatgagt atgaagctca tcaagagaga gtattagcca ggatcaacaa
     1081 gcacaataat cagcaagcac tcagtcacag cattgaggaa ggactgaaga tacaggccat
     1141 gaacagactg cagcgaggca agaaacaaca gattgaaaat ggtagtggag cagaagataa
     1201 tggtgacagt tcacactgca gtaatgcatc cacacatagc aatcaggaag caggccctag
     1261 taacaaacgg accaaaacat ctgatgattc tgggctagag cttgataata acaatgcagc
     1321 aatggcaatt gatccagtaa tggatggtgc tagtgaaatt gaattagtat tcaggcctca
     1381 tcccacactt atggaaaaag atgacagtgc acagacgaga tacataaaga cttctggtaa
     1441 cgccactgtt gatcacttat ccaagtatct ggctgtgagg ttagctttag aagaacttcg
     1501 aagcaaaggt gaatcaaacc agatgaacct tgatacagcc agtgagaagc agtataccat
     1561 ttatatagca acagccagtg gccagttcac tgtattaaat ggctcttttt ctttggaatt
     1621 ggtcagtgag aaatactgga aagtgaacaa acccatggaa ctttattacg cacctacaaa
     1681 ggagcacaaa ctcgagtacc catacgacgt acctgattat gcaacttatc cttatgatgt
     1741 acctgattat gctagctatc cttatgatgt cccagactac gcatagtgag ggcccgatat
     1801 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1861 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1921 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1981 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2041 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2101 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2161 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     2221 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2281 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2341 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2401 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2461 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     2521 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     2581 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     2641 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     2701 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2761 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2821 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     2881 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2941 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3001 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3061 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3121 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3361 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3421 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3481 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3541 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3601 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3661 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3721 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3781 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3841 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3901 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3961 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4021 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4081 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4141 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4201 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4261 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4321 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac 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 tctcaatgct 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 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5581 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5641 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5701 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6841 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     6901 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6961 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7021 atctgcttag g
//
