LOCUS       V042798                 6998 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042798
VERSION     V042798
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             911..931
                     /codon_start=1
                     /locus_tag=""
                     /note=""
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /protein_id=""
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             932..952
                     /codon_start=1
                     /locus_tag=""
                     /note=""
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /protein_id=""
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             953..973
                     /codon_start=1
                     /locus_tag=""
                     /note=""
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /protein_id=""
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             974..2713
                     /label="METTL3(NM_019852)"
                     /note="METTL3(NM_019852)"
                     /gene="METTL3"
     polyA_signal    2760..2984
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3030..3458
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3472..3801
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3868..4467
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4641..4762
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4811..4827)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4835..4851
                     /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(4859..4889)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4904..4925
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5213..5798)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5969..6829)
                     /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(6830..6934)
                     /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 cgccaccatg ccaaaaaaga agagaaaggt tccaaaaaag aagagaaagg tcccaaaaaa
      961 gaagagaaag gtaatgtcgg acacgtggag ctctatccag gcccacaaga agcagctgga
     1021 ctctctgcgg gagaggctgc agcggaggcg gaagcaggac tcggggcact tggatctacg
     1081 gaatccagag gcagcattgt ctccaacctt ccgtagtgac agcccagtgc ctactgcacc
     1141 cacctctggt ggccctaagc ccagcacagc ttcagcagtt cctgaattag ctacagatcc
     1201 tgagttagag aagaagttgc tacaccacct ctctgatctg gccttaacat tgcccactga
     1261 tgctgtgtcc atctgtcttg ccatctccac gccagatgct cctgccactc aagatggggt
     1321 agaaagcctc ctgcagaagt ttgcagctca ggagttgatt gaggtaaagc gaggtctcct
     1381 acaagatgat gcacatccta ctcttgtaac ctatgctgac cattccaagc tctctgccat
     1441 gatgggtgct gtggcagaaa agaagggccc tggggaggta gcagggactg tcacagggca
     1501 gaagcggcgt gcagaacagg actcgactac agtagctgcc tttgccagtt cgttagtctc
     1561 tggtctgaac tcttcagcat cggaaccagc aaaggagcca gccaagaaat caaggaaaca
     1621 tgctgcctca gatgttgatc tggagataga gagccttctg aaccaacagt ccactaagga
     1681 acaacagagc aagaaggtca gtcaggagat cctagagcta ttaaatacta caacagccaa
     1741 ggaacaatcc attgttgaaa aatttcgctc tcgaggtcgg gcccaagtgc aagaattctg
     1801 tgactatgga accaaggagg agtgcatgaa agccagtgat gctgatcgac cctgtcgcaa
     1861 gctgcacttc agacgaatta tcaataaaca cactgatgag tctttaggtg actgctcttt
     1921 ccttaataca tgtttccaca tggatacctg caagtatgtt cactatgaaa ttgatgcttg
     1981 catggattct gaggcccctg gcagcaaaga ccacacgcca agccaggagc ttgctcttac
     2041 acagagtgtc ggaggtgatt ccagtgcaga ccgactcttc ccacctcagt ggatctgttg
     2101 tgatatccgc tacctggacg tcagtatctt gggcaagttt gcagttgtga tggctgaccc
     2161 accctgggat attcacatgg aactgcccta tgggaccctg acagatgatg agatgcgcag
     2221 gctcaacata cccgtactac aggatgatgg ctttctcttc ctctgggtca caggcagggc
     2281 catggagttg gggagagaat gtctaaacct ctgggggtat gaacgggtag atgaaattat
     2341 ttgggtgaag acaaatcaac tgcaacgcat cattcggaca ggccgtacag gtcactggtt
     2401 gaaccatggg aaggaacact gcttggttgg tgtcaaagga aatccccaag gcttcaacca
     2461 gggtctggat tgtgatgtga tcgtagctga ggttcgttcc accagtcata aaccagatga
     2521 aatctatggc atgattgaaa gactatctcc tggcactcgc aagattgagt tatttggacg
     2581 accacacaat gtgcaaccca actggatcac ccttggaaac caactggatg ggatccacct
     2641 actagaccca gatgtggttg cacggttcaa gcaaaggtac ccagatggta tcatctctaa
     2701 acctaagaat ttatagctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2761 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2821 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2881 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2941 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3001 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3061 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3481 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3721 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3781 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3841 caagagacag gatgaggatc gtttcgcatg accgagtaca agcccacggt gcgcctcgcc
     3901 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3961 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4021 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4081 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4141 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4201 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4261 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     4321 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4381 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4441 tgcatgaccc gcaagcccgg tgcctgagcg ggactctggg gttcgaaatg accgaccaag
     4501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4621 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4741 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6961 acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
