LOCUS       V023352                 7260 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023352
VERSION     V023352
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1940
                     /label="Pigk(NM_001011953)"
                     /note="Pigk(NM_001011953)"
                     /gene="Pigk"
     CDS             1947..2012
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2036..2624
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2690..2738
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2940..3368
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3382..3711
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3778..4572
                     /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    4748..4869
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4918..4934)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4942..4958
                     /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(4966..4996)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5011..5032
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5320..5908)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6079..6939)
                     /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(6940..7044)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatccg ccaccatggc gtccccctgc ttctttaccc
      781 tgccagtggc tgtcttggcc accttggcgt tcttgtctct cggcagctcc gccgctagtc
      841 acatcgagga tcaagcagaa caattcttta gaagtggaca tacaaataac tgggctgttt
      901 tggtatgcac atcccgattc tggtttaatt accgacatgt tgcaaatact ctctctgttt
      961 acagaagcgt caagaggcta ggtattccag acagtcacat tgtccttatg cttgctgatg
     1021 acatggcgtg caatgctcgg aaccccaagc cagccacagt gttcagccac aagaacatgg
     1081 agctcaatgt gtatggggac gacgtggaag tggactacag aagctatgag gtgaccgtgg
     1141 agaacttttt acgggtatta actgggaggg ttccacccag cacccctcgt tcaaagcgtc
     1201 ttctctccga cgacagaagc aatatcctca tttatatgac aggtcatgga ggaaacggtt
     1261 ttttgaaatt ccaagattct gaagaaatca ctaacataga acttgcagat gcgtttgaac
     1321 agatgtggca gaaaagacgc tacaacgagc tgctgttcat tatcgacact tgccagggcg
     1381 cgtccatgta tgagcggttt tactctccta acatcatggc cttggcgagt agccaagtgg
     1441 gcgaggattc cctgtcgcac cagcccgacc ccgcaattgg cgtccacctt atggacaggt
     1501 acacattcta tgtcttggaa tttttggaag agattaatcc agctagccaa actaacatga
     1561 atgacctttt tcaagtgtgt cccaaaagcc tctgtgtgtc tacccctgga catcgtactg
     1621 accttttcca gcgagatcct aaaaaggtcc tgataaccga tttcttcgga agcgtgcgga
     1681 aggtggaaat cacgacagag acgatcagtt tgcagtggga ttcacaagtc gtggacagca
     1741 gttctgccga ggacggaatg gcggaggagc ccacggaacc tctcaagttc gctgagcagc
     1801 tcccggtggc tcagataata caccagaagc cgaagctgaa ggactggcac cctcccggag
     1861 gcttcatcct ggggctgtgg gcgctcatca tcatggtctt ctttaagacc tacggcatca
     1921 agcacatgaa gttcattttt ctcgaggact acaaagacca tgacggtgat tataaagatc
     1981 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2401 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2641 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2701 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2761 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2821 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     2881 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     2941 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3001 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3061 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3121 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3181 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3241 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3301 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3361 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3541 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3601 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3661 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3721 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3781 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3841 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3901 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3961 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4021 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4081 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4141 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4201 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4261 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4321 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4381 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4441 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4501 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4561 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4621 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4681 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4741 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4801 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4861 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7081 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7141 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7201 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
