LOCUS       V019517                 7190 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019517
VERSION     V019517
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..1870
                     /label="WTAP(NM_001270531)"
                     /note="WTAP(NM_001270531)"
                     /gene="WTAP"
     CDS             1877..1942
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1966..2554
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2620..2668
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2870..3298
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3312..3641
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3708..4502
                     /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    4678..4799
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4848..4864)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4872..4888
                     /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(4896..4926)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4941..4962
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5250..5838)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6009..6869)
                     /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(6870..6974)
                     /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 ccatgaccaa cgaagaacct cttcccaaga aggttcgatt
      721 gagtgaaaca gacttcaaag ttatggcaag agatgagtta attctaagat ggaaacaata
      781 tgaagcatat gtacaagctt tggagggcaa gtacacagat cttaactcta atgatgtaac
      841 tggcctaaga gagtctgaag aaaaactaaa gcaacaacag caggagtctg cacgcaggga
      901 aaacatcctt gtaatgcgac tagcaaccaa ggaacaagag atgcaagagt gtactactca
      961 aatccagtac ctcaagcaag tccagcagcc gagcgttgcc caactgagat caacaatggt
     1021 agacccagcg atcaacttgt ttttcctaaa aatgaaaggt gaactggaac agactaaaga
     1081 caaactggaa caagcccaaa atgaactgag tgcctggaag tttacgcctg atagccaaac
     1141 agggaaaaag ttaatggcga agtgtcgaat gcttatccag gagaatcaag agcttggaag
     1201 gcagctgtcc cagggacgta ttgcacaact tgaagcagag ttggctttac agaagaaata
     1261 cagtgaggag cttaaaagca gtcaggatga actgaatgac ttcatcatcc agcttgatga
     1321 agaagtagag ggtatgcaga gtaccattct agttctgcag cagcagctga aggagacacg
     1381 ccagcagttg gctcagtacc agcagcagca gtctcaggcc tctgccccaa gtaccagcag
     1441 gactacagct tctgaacctg tagaacagtc agaggccaca agtaaagact gcagtcgtct
     1501 gacaaacgga ccaagtaatg gtagctcctc ccgccagagg acgtctgggt ctggatttca
     1561 cagggagggc aacacaaccg aagatgactt tccttcttct ccagggaatg gtaataagtc
     1621 ctccaacagc tcagaggaga gaactggcag aggaggtagt ggttacgtaa atcaactcag
     1681 tgcggggtat gaaagtgtag actctcccac gggcagtgaa aactctctca cacaccaatc
     1741 aaatgacaca gactccagtc atgaccctca agaggagaaa gcagtgagtg ggaaaggtaa
     1801 tcgaactgtg ggttcccgcc acgttcagaa tggcttggac tcaagtgtaa atgtacaggg
     1861 ttcagttttg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     1921 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat
     1981 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2041 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2101 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2161 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2221 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2281 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2341 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2401 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2461 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2521 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     2581 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2641 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2701 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2761 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     2821 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     2881 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2941 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3001 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     3061 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3121 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3181 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3241 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3301 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3361 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3481 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3541 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3601 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3661 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3721 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3781 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3841 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3901 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3961 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4021 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4081 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4141 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4201 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4261 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4321 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4381 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4441 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4501 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4561 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4621 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4801 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4861 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4921 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4981 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5041 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5101 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5161 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5221 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5281 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5341 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5401 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5461 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5521 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5581 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5641 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5701 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5821 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5881 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5941 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6001 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6121 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6181 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6241 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6301 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6361 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6421 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6481 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6541 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6601 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6661 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6721 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6781 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6841 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6901 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6961 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7021 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     7081 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7141 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
