LOCUS       V046220                 9115 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046220
VERSION     V046220
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             706..3774
                     /label="CDCA2(NM_152562)"
                     /note="CDCA2(NM_152562)"
                     /gene="CDCA2"
     CDS             3781..3807
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3811..3837
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3841..3867
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3891..4479
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4545..4593
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4795..5223
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5237..5566
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5633..6427
                     /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    6603..6724
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6773..6789)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6797..6813
                     /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(6821..6851)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6866..6887
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7175..7763)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7934..8794)
                     /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(8795..8899)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgga tgccaattca
      721 aaagacaagc cccctgaaac caaggagtct gcaatgaata atgctggaaa tgcctctttc
      781 attttgggaa ctgggaagat tgtgactcct cagaagcatg ccgaattacc tcctaatcct
      841 tgcacaccag atacttttaa atcacctttg aacttttcca cagtaaccgt agagcaattg
      901 ggaattacac ctgaaagctt tgttaggaac tctgcaggaa agtcatcatc ctaccttaaa
      961 aaatgtagac gacgttctgc agtcggtgct cggggctctc ctgaaacaaa ccatctgatt
     1021 cgtttcattg ctcggcagca aaatataaag aatgctagga aatctccttt ggcacaagat
     1081 tctccttccc agggcagccc tgcactgtat cgaaatgtta acactttaag agaacgaata
     1141 tcagccttcc agtcagcttt tcactccata aaggaaaacg agaaaatgac cggctgtctg
     1201 gaattctcag aggcaggaaa agagtccgag atgacagact tgaccagaaa ggaaggtctc
     1261 agcgcttgcc agcagtctgg gttccctgca gtgttgtcct ccaaacgtcg gagaatatcc
     1321 tatcagagag actctgatga aaatctgacg gatgctgaag gaaaagtaat tggtctccag
     1381 atattcaata ttgatacaga cagagcatgt gcagttgaaa cttctgtaga tctttctgag
     1441 atatcatcta aacttggttc aacacagtct ggatttttag ttgaagagtc tcttcccctt
     1501 tcagagctca cagagacttc aaatgcacta aaggttgctg actgtgtagt gggcaaagga
     1561 tcaagtgatg ccgtttcgcc tgacacgttc acagcagaag tgagctcaga cgcagtccct
     1621 gatgtcaggt caccagctac tccagcctgc aggagggacc ttcccacccc caagaccttt
     1681 gtacttcgtt ctgtactgaa gaaaccctct gttaagatgt gtctagagag cttacaggaa
     1741 cactgtaaca acctctatga tgatgatggg actcatccga gcttaatctc aaatctccca
     1801 aactgttgca aagagaaaga agcagaagat gaagaaaatt ttgaagcacc tgcctttcta
     1861 aatatgagga agaggaagag agttactttt ggagaggact taagcccgga agtgtttgat
     1921 gaatctttgc cagcaaatac tccattgcgt aaaggaggaa cacctgtttg taaaaaagac
     1981 ttcagtggtc tcagttccct gctgcttgag cagtcacctg ttcctgagcc attacctcaa
     2041 ccagattttg atgacaaggg ggagaatctt gaaaacatag aaccacttca agtatcattt
     2101 gccgttctca gttctcctaa taaatcatca atctctgaga ccctttcagg cactgatacc
     2161 tttagttctt caaataacca tgagaaaata tcctctccta aagttggtag aataacaagg
     2221 acttctaaca gaagaaatca attggtcagt gttgtagaag agagtgtttg caacttattg
     2281 aatacagaag ttcagccttg taaagaaaag aaaattaata ggaggaagtc tcaagaaaca
     2341 aagtgtacaa agagagcact tcctaagaag agtcaggttt taaaaagttg cagaaagaag
     2401 aaaggaaagg gaaagaaaag tgttcagaaa tctttatatg gggaaagaga cattgcttct
     2461 aagaagcccc tcctcagtcc tattcccgag ctgcctgaag tccctgagat gacaccttcc
     2521 attccgagca tccgaagact gggttcaggt tatttcagtt caaatggcaa actggaagaa
     2581 gtgaagactc ctaaaaatcc agtgaaaaga aaggatcttt tgcgtcatga cccagatttg
     2641 catatgcatc aaggctatga taaatatgat gtctctgaat tctgctctta tataaaaagt
     2701 tcctcatcgc ttggcaatgc tacttctgat gaagatccaa atacaaatat aatgaacatt
     2761 aatgaaaata aaaatattcc aaaagcaaaa aataagtcag aaagtgaaaa tgaaccaaaa
     2821 gctggaactg acagtcctgt ttcttgtgct tctgtaactg aagaacgtgt ggcatcagat
     2881 agtcccaaac ctgctctgac cctgcagcag ggtcaagaat tttctgctgg tggtcaaaat
     2941 gcagaaaacc tttgtcagtt ctttaaaatt tcaccagatt taaacataaa gtgtgaaaga
     3001 aaggatgact tcttaggagc tgcagaagga aaactgcaat gcaatcgttt aatgcctaat
     3061 tcacaaaaag actgtcattg tttaggagat gtcttaattg aaaatacgaa agaatctaaa
     3121 agccagagtg aggatttggg aagaaaaccc atggaaagta gcagtgttgt gagttgcaga
     3181 gacaggaaag atagaagacg ttccatgtgt tattctgatg gtcgaagttt acatttggaa
     3241 aaaaatggaa atcacacacc atcctccagt gtgggcagct ctgtagaaat tagtttagaa
     3301 aattctgaac tgtttaaaga tttgtctgat gccattgagc aaacctttca gaggagaaat
     3361 agtgaaacca aagtgcgacg tagcacgagg ctacagaagg atttagaaaa cgaaggtctt
     3421 gtatggattt cacttccact tccttccact tcccaaaaag ccaaaagaag aacaatatgt
     3481 acatttgaca gcagtggatt tgaaagtatg tctcccataa aagaaactgt gtcctccaga
     3541 caaaaaccgc agatggcacc tcccgtctca gatccagaaa acagccaggg ccctgctgct
     3601 ggttcttccg atgaacctgg taagaggagg aagagctttt gtatatctac acttgcaaat
     3661 actaaagcca cttcccagtt caaaggctac cggagaagat cctctcttaa tgggaaggga
     3721 gagagctctc tgactgcctt ggaaaggatt gaacataatg gagaaagaaa gcagctcgag
     3781 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc
     3841 tatccttatg atgtcccaga ctacgcatag tgagggcccg atatctcgac aatcaacctc
     3901 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3961 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4021 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4081 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4141 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4201 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4261 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4321 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4381 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4441 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     4501 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4561 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4621 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4681 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4741 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     4801 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4861 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4921 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4981 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5041 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5101 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5161 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5221 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5281 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5401 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5461 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5521 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5581 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5641 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5701 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5761 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5821 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5881 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5941 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6001 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6061 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6121 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6181 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6241 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6301 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6361 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6421 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     6481 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6541 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6601 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6721 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8041 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8761 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8821 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8881 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8941 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     9001 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     9061 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
