LOCUS       V020306                 8132 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020306
VERSION     V020306
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             893..919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             938..3652
                     /label="CIP2A(NM_020890)"
                     /note="CIP2A(NM_020890)"
                     /gene="CIP2A"
     polyA_signal    3699..3923
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3969..4397
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4411..4740
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4807..5601
                     /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    5775..5896
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5945..5961)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5969..5985
                     /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(5993..6023)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6038..6059
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6347..6932)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7103..7963)
                     /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(7964..8068)
                     /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 tgtaccctta
      901 tgacgtcccc gattacgccg gtaccgagct cggatccatg gactccactg cctgcttgaa
      961 gtccttgctc ctgactgtca gtcagtacaa agccgtgaag tcagaggcga acgccactca
     1021 gcttttgcgg cacttggagg taatttctgg acagaaactc acacgactat ttacatcaaa
     1081 tcagatatta acaagtgaat gcttgagttg ccttgtagag ctacttgaag accccaacat
     1141 aagtgcttca ctgatcttaa gtattatcgg tttgctgtct caactagcag tagacattga
     1201 aaccagagat tgtcttcaga atacatataa tctgaatagt gtgctggcgg gagtggtttg
     1261 tcggagcagc cacactgatt cggtgttttt gcagtgcatt caacttctac agaagttaac
     1321 atataatgtc aaaattttct attctggtgc caatatagat gaattaatta cgttcctgat
     1381 agatcacatt caatcttctg aagatgagtt aaaaatgcct tgtctaggat tattggcaaa
     1441 tctttgtcgg cacaatcttt ctgttcaaac gcacataaag acattgagta atgtgaaatc
     1501 tttttatcga actcttatca ccttgttggc ccatagtagt ttaactgtgg ttgtgtttgc
     1561 actttcaata ttatccagtt tgacattaaa tgaagaggtg ggggaaaagc tattccatgc
     1621 tcgaaacatt catcagactt ttcaactaat atttaatatt ctcataaacg gtgatggcac
     1681 tctaactaga aagtattcag ttgacctact gatggatctc cttaagaatc ctaaaattgc
     1741 tgattatctc accagatatg agcacttttc ttcatgtctt caccaagtat taggtcttct
     1801 taatggaaag gatcctgatt cctcttcaaa ggttttagaa ttacttcttg ccttctgttc
     1861 agtgactcag ctgcgccata tgctcactca gatgatgttt gaacagtctc cacctggcag
     1921 cgccactctg ggaagccata ctaaatgttt agaacctact gtggctctac tgcgctggtt
     1981 aagccaacct ttggacggat cagaaaactg ttctgtttta gcattggagt tgttcaagga
     2041 aatatttgag gatgtcatag atgctgctaa ctgttcctcg gctgatcgtt ttgtgaccct
     2101 tctgctgcct acaatccttg atcaacttca gttcacagaa caaaatctag atgaggcttt
     2161 aacaagaaaa aaatgtgaaa ggattgccaa ggccattgaa gttttgttaa ctctctgtgg
     2221 agatgataca ctaaaaatgc atattgcaaa aatcttgaca actgtcaagt gtaccactct
     2281 tatagaacaa caatttacat atggcaagat tgacctggga tttggaacaa aggttgcaga
     2341 ttctgaatta tgcaaacttg ctgctgatgt aattttgaaa actcttgatt tgattaacaa
     2401 acttaaacca ttggttcctg gtatggaagt aagcttctac aaaatacttc aggacccacg
     2461 tttgattact cctttggctt ttgctttaac gtcagataat agagaacaag tacagtctgg
     2521 actgagaata ttattggagg ctgctccact gccagatttt cctgctttag tacttggaga
     2581 aagtatagca gcaaacaatg cctatagaca acaggaaaca gaacatatac ccagaaaaat
     2641 gccctggcaa tcatcaaatc acagttttcc aacatcaata aagtgtttaa ctcctcattt
     2701 gaaagatggt gttcctggat tgaatattga agaattaata gagaaacttc agtctggaat
     2761 ggtggtaaag gatcagattt gtgatgtgag aatatctgac ataatggatg tatatgaaat
     2821 gaaactatcc acattagctt ccaaagaaag caggctacaa gatcttttgg aaacaaaagc
     2881 tctagccctt gcacaggctg atagactgat tgctcagcat cgctgtcaaa gaactcaagc
     2941 tgaaacagag gcacggacac ttgctagtat gttgagagaa gttgagagaa aaaatgaaga
     3001 gcttagtgtg ttgctgaagg cgcagcaagt tgaatcagaa agagcgcaga gtgatattga
     3061 gcatctcttt caacataata ggaagttaga gtctgtggct gaagaacatg aaatactgac
     3121 aaaatcctac atggaacttc ttcagagaaa tgaaagtact gaaaagaaga ataaagattt
     3181 acagatcaca tgtgattctc tgaataaaca aattgagaca gtgaaaaagt tgaatgagtc
     3241 actcaaggaa caaaatgaaa aaagtattgc ccaattaata gagaaagaag aacagagaaa
     3301 agaagtacag aatcagctag tagacagaga acataagcta gcaaatttgc atcaaaaaac
     3361 aaaagtacaa gaagaaaaga ttaaaacctt acaaaaggaa agggaagata aggaagaaac
     3421 cattgatatc cttagaaaag aattaagcag aacagaacag ataagaaaag agttgagcat
     3481 taaggcttcc tccctagagg ttcaaaaggc acaattagaa ggtcgtttgg aagagaaaga
     3541 gtccttggtg aaacttcagc aagaggaatt gaacaaacac tcccacatga tagcaatgat
     3601 ccacagttta agtggtggaa aaataaatcc agaaactgtg aatctcagta tatagctcga
     3661 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3721 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3781 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3841 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3901 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3961 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4021 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4081 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4141 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4201 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4261 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4321 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4381 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4621 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4681 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4741 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4801 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4861 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4921 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4981 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5041 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5101 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5161 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5221 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5281 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5341 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5401 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5461 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5521 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5581 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5641 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5701 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5761 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5821 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5881 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5941 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6001 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6061 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6121 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6181 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6241 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6301 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6361 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6421 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6481 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6541 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6601 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6661 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6721 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6781 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6841 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6961 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7021 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7081 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7141 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7201 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7261 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7321 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7381 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7441 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7501 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7561 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7621 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7681 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7741 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7801 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7861 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7921 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7981 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8041 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8101 acggatcggg agatctcccg atcccctatg gt
//
