LOCUS       V020101                 6890 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020101
VERSION     V020101
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..1546
                     /label="OTUD6A(NM_207320)"
                     /note="OTUD6A(NM_207320)"
                     /gene="OTUD6A"
     CDS             1553..1642
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1666..2254
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2320..2368
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2570..2998
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3012..3341
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3408..4202
                     /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    4378..4499
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4548..4564)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4572..4588
                     /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(4596..4626)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4641..4662
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4950..5538)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5709..6569)
                     /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(6570..6674)
                     /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 ccatggatga tccgaagagt gaacagcagc gcatactgcg
      721 ccgccaccaa cgcgagaggc aggagctgca ggcccagatc cggagcttaa aaaactcggt
      781 ccccaagacc gacaagacga aaagaaagca gttgctccaa gacgtggccc gcatggaggc
      841 cgagatggct cagaagcacc ggcaggagct ggagaagttc caagacgaca gtagcattga
      901 atctgtcgtc gaagacctgg ccaagatgaa tctggaaaac cggcctcccc gctcctccaa
      961 agcccacaga aagagagaaa gaatggagtc cgaggagagg gagcgccagg agagcatctt
     1021 ccaggctgag atgtcggagc acctggccgg cttcaagcgc gaggaggagg agaagctcgc
     1081 cgccatcctg ggagccaggg gtctggagat gaaagcgatc ccggccgacg gccactgcat
     1141 gtaccgcgcc atccaagacc agctggtgtt cagcgtgtct gtggagatgc tgcgctgccg
     1201 caccgccagc tacatgaaga agcacgtcga cgagttcctg cccttcttca gcaaccccga
     1261 gaccagcgac tccttcggct acgacgactt catgatctac tgcgacaaca tcgtgcgcac
     1321 cacggcatgg ggaggccagc tggagctgag ggccctgtcg cacgtcctga agacccccat
     1381 cgaggtgatc caggccgact cgcccacctt gatcatcggg gaggagtacg tcaagaagcc
     1441 gatcatcctg gtctacctgc gctatgccta cagcctcggc gagcactaca actccgtgac
     1501 accgctcgag gccggcgccg ccgggggcgt gctcccgcgt ctcctgctcg aggaacaaaa
     1561 actaatatca gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa
     1621 gttaatttcc gaggaggact tatagtgagg gcccgatatc tcgacaatca acctctggat
     1681 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1741 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1801 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     1861 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     1921 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     1981 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2041 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2101 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2161 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2221 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     2281 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2341 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2401 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2461 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     2521 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     2581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2641 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2701 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     2761 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2881 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2941 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3001 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3181 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3241 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3301 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3361 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3421 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3481 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3541 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3601 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3661 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3721 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3781 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3841 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3901 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3961 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4021 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4081 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4141 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4201 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4261 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4321 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4381 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4441 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4501 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6721 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     6781 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     6841 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
