LOCUS       V024444                 6925 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024444
VERSION     V024444
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             937..2676
                     /label="METTL3(NM_019852)"
                     /note="METTL3(NM_019852)"
                     /gene="METTL3"
     CDS             2689..2754
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2789..3013
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3059..3487
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3501..3830
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3842..4441
                     /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    4601..4722
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4771..4787)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4795..4811
                     /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(4819..4849)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4864..4885
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5173..5758)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5929..6789)
                     /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(6790..6894)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcga tatcggatcc gccaccatgt cggacacgtg gagctctatc
      961 caggcccaca agaagcagct ggactctctg cgggagaggc tgcagcggag gcggaagcag
     1021 gactcggggc acttggatct acggaatcca gaggcagcat tgtctccaac cttccgtagt
     1081 gacagcccag tgcctactgc acccacctct ggtggcccta agcccagcac agcttcagca
     1141 gttcctgaat tagctacaga tcctgagtta gagaagaagt tgctacacca cctctctgat
     1201 ctggccttaa cattgcccac tgatgctgtg tccatctgtc ttgccatctc cacgccagat
     1261 gctcctgcca ctcaagatgg ggtagaaagc ctcctgcaga agtttgcagc tcaggagttg
     1321 attgaggtaa agcgaggtct cctacaagat gatgcacatc ctactcttgt aacctatgct
     1381 gaccattcca agctctctgc catgatgggt gctgtggcag aaaagaaggg ccctggggag
     1441 gtagcaggga ctgtcacagg gcagaagcgg cgtgcagaac aggactcgac tacagtagct
     1501 gcctttgcca gttcgttagt ctctggtctg aactcttcag catcggaacc agcaaaggag
     1561 ccagccaaga aatcaaggaa acatgctgcc tcagatgttg atctggagat agagagcctt
     1621 ctgaaccaac agtccactaa ggaacaacag agcaagaagg tcagtcagga gatcctagag
     1681 ctattaaata ctacaacagc caaggaacaa tccattgttg aaaaatttcg ctctcgaggt
     1741 cgggcccaag tgcaagaatt ctgtgactat ggaaccaagg aggagtgcat gaaagccagt
     1801 gatgctgatc gaccctgtcg caagctgcac ttcagacgaa ttatcaataa acacactgat
     1861 gagtctttag gtgactgctc tttccttaat acatgtttcc acatggatac ctgcaagtat
     1921 gttcactatg aaattgatgc ttgcatggat tctgaggccc ctggcagcaa agaccacacg
     1981 ccaagccagg agcttgctct tacacagagt gtcggaggtg attccagtgc agaccgactc
     2041 ttcccacctc agtggatctg ttgtgatatc cgctacctgg acgtcagtat cttgggcaag
     2101 tttgcagttg tgatggctga cccaccctgg gatattcaca tggaactgcc ctatgggacc
     2161 ctgacagatg atgagatgcg caggctcaac atacccgtac tacaggatga tggctttctc
     2221 ttcctctggg tcacaggcag ggccatggag ttggggagag aatgtctaaa cctctggggg
     2281 tatgaacggg tagatgaaat tatttgggtg aagacaaatc aactgcaacg catcattcgg
     2341 acaggccgta caggtcactg gttgaaccat gggaaggaac actgcttggt tggtgtcaaa
     2401 ggaaatcccc aaggcttcaa ccagggtctg gattgtgatg tgatcgtagc tgaggttcgt
     2461 tccaccagtc ataaaccaga tgaaatctat ggcatgattg aaagactatc tcctggcact
     2521 cgcaagattg agttatttgg acgaccacac aatgtgcaac ccaactggat cacccttgga
     2581 aaccaactgg atgggatcca cctactagac ccagatgtgg ttgcacggtt caagcaaagg
     2641 tacccagatg gtatcatctc taaacctaag aatttactcg agaccggtga ctacaaagac
     2701 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagtgaccg
     2761 gtaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2821 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2881 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2941 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3001 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3061 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3121 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3181 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3241 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3301 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3361 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3421 gggattttgg ccatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3481 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3661 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3721 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3781 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3841 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3901 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3961 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4021 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4081 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4141 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4201 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4261 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4321 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4381 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4441 attcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4501 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4561 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4621 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4681 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4741 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4801 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4861 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4921 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4981 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5041 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5581 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5701 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6901 tccccgaaaa gtgccacctg acgtc
//
