LOCUS       V029525                 7705 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029525
VERSION     V029525
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3144
                     /label="Ythdc1(NM_133423)"
                     /note="Ythdc1(NM_133423)"
                     /gene="Ythdc1"
     CDS             3151..3177
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3181..3207
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3211..3237
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3272..3496
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3542..3970
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3984..4313
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4380..5174
                     /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    5348..5469
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5518..5534)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5542..5558
                     /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(5566..5596)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5611..5632
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5920..6505)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6676..7536)
                     /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(7537..7641)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcggccg acagccggga ggagaaagat
      961 ggggaactta atgttttaga tgatattttg actgaagtac cagaacagga tgatgaactg
     1021 tataatccag agagtgaaca agataaaaat gagaaaaaag gatcaaaaag aaaaagtgaa
     1081 agaatggaat ctattgacac caagcgacag aagccttcta tccattcaag acaactgatt
     1141 tctaagccac taagctcatc tgttagcaat aataaaagaa tagttagtac aaaaggaaag
     1201 tcggttacag aatataaaaa tgaggaatat caaagatctg aaaggaacaa gcgtctagat
     1261 gctgatcgaa aaattcgtct gtcaagcagt tcctctagag aaccttacaa gagtcaacca
     1321 gaaaaacctt gtctacggaa aagggattct gaaagaaggg ccaagtctcc tacaccagat
     1381 ggttctgaga gaattgggct tgaagttgat agacgtgcaa gcagatccag ccagtcttca
     1441 aaggaagagg ggaactctga ggagtatggc tctgaccacg agacaggaag cagtgcttct
     1501 tctgaacagg gcaacaacac tgagaatgag gaggaaggag gggaagaaga tgtagaggaa
     1561 gatgaagagg tagatgaaga tggagatgat gatgaggaag tggatgaaga tgcggaggag
     1621 gaggaggacg aggaagaaga tgaggaggag gaggatgagg aagaagaaga ggaagaggaa
     1681 gaagaatatg aacaggatga gagagatcag aaggaagaag ggaatgatta tgacacccgt
     1741 agtgaggcca gtgattctgg ttctgagtct gtttccttca cagatggatc tgtcaggtct
     1801 ggttcaggaa cagatggatc agatgagaaa aagaaggaaa ggaagagagc tcgaggcata
     1861 tcacccattg tctttgatag aagtggcagt tctgcatcag agtcatatgc aggttcagaa
     1921 aagaagcatg agaaattatc atcttccgtt cgtgctgtcc gaaaagatca aaccagtaaa
     1981 ctcaaatctg tccttcagga tgcaagattt ttcctcataa agagtaacaa ccatgagaat
     2041 gtgtctcttg ccaaagcaaa gggtgtatgg tccacattac ctgtaaatga gaagaaatta
     2101 aatcttgcgt ttagatctgc aaggagtgtt atattaatat tttctgtcag ggaaagtgga
     2161 aagtttcaag gttttgccag attgtcatca gaatcgcatc atggtggctc tcctatacat
     2221 tgggtgcttc cagcaggaat gagtgctaaa atgcttggag gtgtttttaa aattgactgg
     2281 atttgcaggc gtgaattacc ctttactaag tcagctcatc tcaccaatcc ctggaatgaa
     2341 cataagccag taaagattgg acgtgatgga caggaaattg aacttgaatg tggaacccag
     2401 ctttgtcttc tgtttccccc tgatgaaagt attgacttgt atcagctcat tcataaaatg
     2461 cgtcacaaga gaagaatgca ttctcagcct cgatcaagag gacgtccatc ccgtcgagaa
     2521 ccagtccggg atgtgggaag gcgtcgacca gaagattatg atattcataa cagcagaaag
     2581 aaaccaagga ttgactatcc ccctgagttt caccagagac cagggtattt aaaggatccc
     2641 cgataccagg aagttgacag tttcacaaat cttattccca acagacgatt ttcaggagtt
     2701 cgccgagatg tgtttttaaa tgggtcctac aatgattatg tgagggaatt tcataacatg
     2761 ggaccaccgc ctccttggca aggaatgcct ccttacccgg gaatagaaca gcctccacac
     2821 catccctact accagcacca tgccccgcct cctcaagccc acccccctta ctcaggacac
     2881 catccggtac cacatgaagc aagatacaga gataaacgag tacatgacta tgatatgagg
     2941 gttgatgatt tccttcgccg cacacaagcc gttgtcagtg gtcggagaag tagaccccga
     3001 gaaagagatc gggagcgaga gcgagaccgc cctagagata acagaagaga tagagagcga
     3061 gacagaggtc gtgatcgaga aagagagaga gaaagaatat gtgatcggga cagagaccga
     3121 ggggaaagag gtcgttatcg aagactcgag tacccatacg acgtacctga ttatgcaact
     3181 tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga ctacgcatga
     3241 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3301 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3361 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3421 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3481 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3541 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3961 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4201 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4261 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4321 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4381 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4441 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4501 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4561 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4621 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4681 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4741 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4801 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4861 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4921 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4981 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5041 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5101 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5161 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5221 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5281 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5341 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5461 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5521 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5581 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5641 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6361 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7681 gggagatctc ccgatcccct atggt
//
