LOCUS       V020180                 6620 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020180
VERSION     V020180
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..1300
                     /label="MSC(NM_005098)"
                     /note="MSC(NM_005098)"
                     /gene="MSC"
     CDS             1307..1372
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1396..1984
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2050..2098
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2300..2728
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2742..3071
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3138..3932
                     /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    4108..4229
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4278..4294)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4302..4318
                     /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(4326..4356)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4371..4392
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4680..5268)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5439..6299)
                     /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(6300..6404)
                     /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 ccatgtccac gggctcggtg agtgatccgg aggagatgga
      721 gcttcggggg ctgcagcggg agtacccggt ccccgcctcc aagaggccgc ccctccgcgg
      781 cgtagagcgc agctacgcct cgcccagtga caactcgtcg gcagaggagg aggaccccga
      841 cggcgaggag gagcgctgcg ctctgggcac agccggcagc gcggaaggct gcaagaggaa
      901 gcggccccgt gtggctgggg gcggcggcgc aggtggtagc gcgggcggtg gtggcaagaa
      961 gcccctcccg gccaagggct cagccgcaga gtgcaagcag tcgcagcgga acgcggccaa
     1021 cgcccgtgag cgtgcccgga tgcgcgtgct gagcaaagcc ttctccaggc tcaagaccag
     1081 cctgccctgg gtgccccccg acactaagct ctccaagctg gacacgctcc ggctggcttc
     1141 cagttacatc gctcacctgc ggcagctgtt gcaggaggac cgctatgaga acggctacgt
     1201 gcacccagtg aacctgacat ggccattcgt ggtctcggga agaccggact ctgacaccaa
     1261 agaagtttcc gcagccaaca gactatgtgg aaccaccgct ctcgaggact acaaagacca
     1321 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     1381 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1441 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1501 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     1561 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     1621 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     1681 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1741 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     1801 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     1861 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     1921 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     1981 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     2041 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     2101 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     2161 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     2221 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     2281 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2341 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2401 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     2461 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2521 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2581 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2641 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     2701 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2821 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2941 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3001 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3061 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3121 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3181 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3241 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3301 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3361 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3421 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3481 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3541 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3601 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3661 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3721 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3781 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3841 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3901 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     3961 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4021 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4081 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4201 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4261 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4321 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4381 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4441 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4861 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5101 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6421 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     6481 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     6541 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     6601 gcatgaagaa tctgcttagg
//
