LOCUS       V019454                 7667 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019454
VERSION     V019454
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             671..2347
                     /label="YTHDF1(NM_017798)"
                     /note="YTHDF1(NM_017798)"
                     /gene="YTHDF1"
     CDS             2354..2419
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2443..3031
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3097..3145
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3347..3775
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3789..4118
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4185..4979
                     /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    5155..5276
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5325..5341)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5349..5365
                     /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(5373..5403)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5418..5439
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5727..6315)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6486..7346)
                     /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(7347..7451)
                     /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 taccgccacc atgtcggcca ccagcgtgga cacccagaga acaaaaggac aagataataa
      721 agtacaaaat ggttcgttac atcagaagga tacagttcat gacaatgact ttgagcccta
      781 ccttactgga cagtcaaatc agagtaacag ttacccctca atgagcgacc cctacctgtc
      841 cagctattac ccgccgtcca ttggatttcc ttactccctc aatgaggctc cgtggtctac
      901 tgcaggggac cctccgattc catacctcac cacctacgga cagctcagta acggagacca
      961 tcattttatg cacgatgctg tttttgggca gcctgggggc ctggggaaca acatctatca
     1021 gcacaggttc aattttttcc ctgaaaaccc tgcgttctca gcatggggga caagtgggtc
     1081 tcaaggtcag cagacccaga gctccgcgta tgggagcagc tacacctacc ccccgagctc
     1141 cctgggtggc acggtggttg atgggcagcc aggctttcac agcgacaccc tcagcaaggc
     1201 ccccgggatg aacagcctgg agcagggcat ggttggcctg aagattgggg acgtcagctc
     1261 ctccgccgtc aagacggtgg gctctgtcgt cagcagcgtg gcactgactg gtgtcctttc
     1321 tggcaacggt gggacaaatg tgaacatgcc agtttcaaag ccgacctcgt gggctgccat
     1381 tgccagcaag cctgcaaaac cacagcctaa aatgaaaaca aagagcgggc ctgtcatggg
     1441 gggtgggctg ccccctccac ccataaagca taacatggac attggcacct gggataacaa
     1501 ggggcctgtg ccgaaggccc cagtccccca gcaggcaccc tctccacagg ctgccccaca
     1561 gccccagcag gtggctcagc ctctcccagc acagccccca gctttggctc aaccgcagta
     1621 tcagagccct cagcagccac cccagacccg ctgggttgcc ccacgcaaca gaaacgcggc
     1681 gtttgggcag agcggagggg ctggcagcga tagcaactct cctggaaacg tccagcctaa
     1741 ttctgccccc agcgtcgaat cccaccccgt ccttgaaaaa ctgaaggctg ctcacagcta
     1801 caacccgaaa gagtttgagt ggaatctgaa aagcgggcgt gtgttcatca tcaagagcta
     1861 ctctgaggac gacatccacc gctccattaa gtactccatc tggtgtagca cagagcacgg
     1921 caacaagcgc ctggacagcg ccttccgctg catgagcagc aaggggcccg tctacctgct
     1981 cttcagcgtc aatgggagtg ggcatttttg tggggtggcc gagatgaagt cccccgtgga
     2041 ctacggcacc agtgccgggg tctggtctca ggacaagtgg aaggggaagt ttgatgtcca
     2101 gtggattttt gttaaggatg tacccaataa ccagctccgg cacatcaggc tggagaataa
     2161 cgacaacaaa ccggtcacaa actcccggga cacccaggag gtgcccttag aaaaagccaa
     2221 gcaagtgctg aaaattatca gttcctacaa gcacacaacc tccatcttcg acgactttgc
     2281 tcactacgag aagcgccagg aggaggagga ggtggtgcgc aaggaacggc agagtcgaaa
     2341 caaacaactc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2401 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     2461 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2521 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2581 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2641 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2701 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2761 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2821 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     2881 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2941 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3001 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     3061 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3121 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3181 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3241 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3301 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     3361 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3421 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3481 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     3541 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3601 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3661 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     3721 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3781 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3961 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4021 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4081 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4141 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4201 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4261 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4321 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4381 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4441 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4501 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4561 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4621 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4681 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4741 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4801 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4861 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4921 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4981 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5281 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7441 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7501 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7561 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7621 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
