LOCUS       V019514                 6590 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019514
VERSION     V019514
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..1246
                     /label="JOSD2(NM_138334)"
                     /note="JOSD2(NM_138334)"
                     /gene="JOSD2"
     CDS             1253..1342
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1366..1954
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2020..2068
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2270..2698
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2712..3041
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3108..3902
                     /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    4078..4199
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4248..4264)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4272..4288
                     /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(4296..4326)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4341..4362
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4650..5238)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5409..6269)
                     /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(6270..6374)
                     /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 ccatgtccca ggccccggga gcacagccga gcccacccac
      721 cgtgtaccac gaacggcagc gcctggagct gtgtgctgtc cacgccctca acaacgttct
      781 gcagcagcag ctctttagcc aggaggctgc cgatgagatc tgcaagaggt tggccccaga
      841 ctcccggctg aaccctcatc gcagcctcct gggcaccggc aactatgatg tcaatgtgat
      901 catggccgct ctgcaggggc tgggcctggc cgccgtgtgg tgggacagga ggaggcccct
      961 gtcccagctg gccctgcccc aggtactggg gctgatcctg aacctgccct cgcccgtgtc
     1021 gctggggctg ctgtcactgc cgctgcgccg gcggcactgg gtggccctgc gccaggtgga
     1081 cggtgtctac tacaacctgg actccaagct gcgggcgccc gaggccctgg gggatgagga
     1141 cggagtcagg gccttcctgg cggctgcgct ggcccagggc ctgtgcgagg tgctgctggt
     1201 agtgaccaag gaggtggagg agaagggcag ctggctgcgg acagacctcg aggaacaaaa
     1261 actaatatca gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa
     1321 gttaatttcc gaggaggact tatagtgagg gcccgatatc tcgacaatca acctctggat
     1381 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1441 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1501 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     1561 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     1621 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     1681 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     1741 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     1801 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     1861 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     1921 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     1981 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2041 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2101 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2161 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     2221 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     2281 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2341 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2401 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     2461 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2521 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2581 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2641 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2701 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2761 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2941 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3001 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3061 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3121 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3181 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3241 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3301 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3361 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3421 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3481 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3541 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3601 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3661 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3721 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3781 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3841 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3901 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3961 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4021 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4081 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4141 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4201 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4261 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4321 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4381 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4441 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     4861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5461 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5521 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5581 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5641 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5701 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5761 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5821 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5881 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5941 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6001 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6061 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6121 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6181 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6241 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6301 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6361 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6421 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     6481 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     6541 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
