LOCUS       V030226                 8674 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030226
VERSION     V030226
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..3333
                     /label="Kpnb1(NM_008379)"
                     /note="Kpnb1(NM_008379)"
                     /gene="Kpnb1"
     CDS             3340..3366
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3370..3396
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3400..3426
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3450..4038
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4104..4152
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4354..4782
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4796..5125
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5192..5986
                     /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    6162..6283
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6332..6348)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6356..6372
                     /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(6380..6410)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6425..6446
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6734..7322)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7493..8353)
                     /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(8354..8458)
                     /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 gctcataacc
      721 atcctcgaga agaccgtgtc tccggatcgg ctggagctgg aagcggcgca gaagttcctc
      781 gagcgtgcgg ccgtggagaa tctgcccacg ttccttgtgg aactgtccag agtgctggca
      841 aacccaggaa acagtcaggt tgccagagtt gcagctggtc tacaaattaa gaactctttg
      901 acatcgaaag atccagatat caaggcacaa taccagcaga ggtggctcgc tattgatgct
      961 aatgctcgac gggaagtcaa gaactatgtt ttgcagacgt tgggcacaga aacgtaccgg
     1021 cctagttcgg cctcacagtg tgtggctggt attgcttgtg cagagatccc agtaagccag
     1081 tggccagagc taattcctca gctggtagcc aatgtcacaa accccaacag cacagagcat
     1141 atgaaagagt ccacattgga agctattggt tacatttgcc aagatataga cccagagcag
     1201 ctacaggata agtccaatga gatcctgact gccataatcc aggggatgag gaaggaggag
     1261 cctagtaaca atgtgaagct ggctgctacc aatgcactcc tgaactcact agagttcacc
     1321 aaagcaaact ttgacaaaga gtctgaaagg cactttatca tgcaagtggt ctgtgaagcc
     1381 acacagtgtc cagacacaag ggtaagagtg gctgctttac agaatctagt gaagataatg
     1441 tccttgtatt accagtacat ggagacatac atgggtcctg ccctttttgc aatcacaatt
     1501 gaagcaatga aaagtgacat tgatgaggtg gctctccaag ggatagagtt ctggtccaat
     1561 gtctgtgatg aggaaatgga tttggccatt gaggcttcag aggcagcaga gcaaggacgc
     1621 cccccggagc acaccagcaa attttacgcc aagggagcac tgcagtactt ggtgcccatc
     1681 ctcacacaga cactgactaa acaggatgaa aacgatgacg acgatgactg gaacccttgc
     1741 aaagcagctg gggtgtgcct catgctcctg tccacctgct gtgaagatga cattgtgccg
     1801 catgtccttc cctttattaa agagcacatc aagaaccctg actggcgata ccgggatgca
     1861 gcagtgatgg cttttggcag tatcttggaa ggaccagagc ctaatcaact gaaaccatta
     1921 gtcatacagg ctatgcccac cctaatagaa ctaatgaaag accccagtgt agttgttcga
     1981 gacacaacag cgtggactgt gggcaggatc tgtgagctgc tgcctgaagc cgccatcaac
     2041 gatgtctacc tggcacccct tttacagtgt ctgattgagg gcctcagtgc tgagcccagg
     2101 gtggcttcaa atgtgtgctg ggctttttcc agtctggctg aagctgcgta tgaagctgca
     2161 gatgtagctg atgatcaaga agaaccagcc acctattgtc tgtcttcttc ctttgaactt
     2221 atagttcaga agctattgga gaccaccgac agacccgatg gacaccagaa taacctgaga
     2281 agctctgcgt atgagtctct catggaaatc gtaaagaaca gtgccaagga ttgttaccct
     2341 gccgtgcaga agaccaccct ggtcattatg gaacggctgc agcaggtgct tcagatggag
     2401 tcccatatcc agagcacatc cgacagaatc cagttcaatg acctccagtc tctactctgc
     2461 gcgactcttc agaatgttct ccggaaagtg cagcatcaag atgctctgca gatctctgat
     2521 gtggtcatgg cctccctgtt aaggatgttc caaagcacag ctgggtctgg gggagtgcaa
     2581 gaagatgccc tgatggcagt tagcacactg gtggaagtgt tgggtggtga attcctcaag
     2641 tacatggagg cctttaaacc attcctgggc attggactga aaaattatgc tgagtaccag
     2701 gtatgtttgg cagctgttgg cttagttgga gacttgtgcc gagccctgca gtctaacatc
     2761 ttgcctttct gtgacgaggt gatgcagctg ctcctggaga acttggggaa tgagaatgtc
     2821 cacaggtctg tgaagccaca gattctgtct gtgtttggtg atattgctct tgccattggt
     2881 ggagagttta aaaaatactt agaggttgta ttgaatactc tacagcaggc ctcccaagcc
     2941 caggttgaca agtcagactt tgacatggtg gattatctga atgagctaag agaaagctgc
     3001 ttggaagctt atacgggaat cgtccaggga ttgaagggag atcaggaaaa cgtacacccg
     3061 gatgtaatgc tggtacagcc cagagtagaa tttattttgt cttttattga tcacattgct
     3121 ggagatgagg atcatacgga cggagtggta gcctgtgctg ctggtctgat aggggacttg
     3181 tgtacagcct tcgggaagga tgtactgaag ttagtagaag ctaggccaat gatccatgaa
     3241 ctattaactg aagggcggag atcgaagact aacaaagcaa agaccctcgc tacgtgggca
     3301 accaaggaac tgaggaaact gaagaaccag gctctcgagt acccatacga cgtacctgat
     3361 tatgcaactt atccttatga tgtacctgat tatgctagct atccttatga tgtcccagac
     3421 tacgcatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     3481 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3541 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3601 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3661 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3721 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3781 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3841 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3901 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3961 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4021 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     4081 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     4141 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4201 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4261 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     4321 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4381 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4441 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4501 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4561 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4621 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4681 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4741 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4801 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4861 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4921 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5041 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5101 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5161 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5221 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5281 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5341 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5401 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5461 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5521 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5581 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5641 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5701 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5761 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5821 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5881 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5941 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6001 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6061 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6121 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6181 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6241 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6301 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6901 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7141 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7501 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7561 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7621 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7681 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7741 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7801 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7861 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7921 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7981 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8041 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8101 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8161 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8221 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8281 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8341 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8401 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8461 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8521 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     8581 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     8641 cttgaccgac aattgcatga agaatctgct tagg
//
