LOCUS       V019501                 7757 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019501
VERSION     V019501
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..2437
                     /label="YTHDF3(NM_152758)"
                     /note="YTHDF3(NM_152758)"
                     /gene="YTHDF3"
     CDS             2444..2509
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2533..3121
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3187..3235
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3437..3865
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3879..4208
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4275..5069
                     /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    5245..5366
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5415..5431)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5439..5455
                     /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(5463..5493)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5508..5529
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5817..6405)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6576..7436)
                     /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(7437..7541)
                     /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 ccatgtcagc cactagcgtg gatcagagac ctaaagggca
      721 aggaaataaa gtttcagtac aaaacggttc gattcatcaa aaagatgctg taaatgatga
      781 tgattttgag ccatacttaa gtagccagac aaatcagagt aacagctatc caccaatgtc
      841 agatccatac atgcctagtt actatgctcc atccattgga tttccatatt ctcttgggga
      901 agcagcgtgg tccacagctg gagaccagcc tatgccatat ctgacaacct atggacaaat
      961 gagtaatgga gaacatcact atataccaga tggtgtattt agtcaacctg gggcattagg
     1021 aaatacccct ccatttcttg gtcaacatgg atttaacttt tttcctggta atgctgattt
     1081 ctctacatgg gggacaagtg gatctcaggg acaatcaaca caaagttctg cttatagtag
     1141 cagttatggc tatccaccta gttctcttgg gagagctatt actgatggac aggctggatt
     1201 tggcaatgat actttgagta aggtgcctgg cattagcagt attgagcaag gcatgactgg
     1261 actgaaaatt ggtggtgacc tgacagctgc agtgacaaaa actgtaggta cagctttgag
     1321 cagcagtggt atgactagca ttgcaaccaa tagtgtgccc ccagttagca gtgcagcacc
     1381 taaaccaacc tcctgggctg ccattgccag aaagcctgcc aaacctcaac cgaaacttaa
     1441 acccaagggc aatgtgggaa ttgggggttc tgctgtacca ccacctccta taaaacacaa
     1501 catgaatatt ggaacttggg atgaaaaagg gtcagtggta aaggctccac caacccaacc
     1561 agttctgcct cctcaaacta taatccagca gcctcagcca ttaattcaac caccaccatt
     1621 ggtgcaaagc caactgcctc aacagcagcc tcaaccacca caaccacagc agcaacaagg
     1681 acctcagcca caggcccagc ctcaccaagt gcagcctcaa cagcagcagc tgcagaatcg
     1741 ctgggtagct cctcgtaaca ggggagcagg cttcaaccag aacaatggag cgggcagtga
     1801 aaactttggt ttaggtgttg tacctgtcag tgcttcacct tctagtgtag aagtgcatcc
     1861 cgtgctggaa aagctaaagg ccataaacaa ctataatccc aaagactttg attggaatct
     1921 gaagaatgga cgtgtgttta taattaaaag ctactctgag gatgacatac atcgttccat
     1981 taaatactct atctggtgta gtactgagca tggtaataag cgtttggatg cagcttaccg
     2041 ttccctgaat gggaaaggcc cactctattt actcttcagt gtgaatggca gtggacattt
     2101 ttgtggagtg gctgaaatga agtctgttgt ggactataat gcgtatgctg gtgtctggtc
     2161 tcaggataag tggaagggca aatttgaagt taaatggatc tttgtcaaag atgttcccaa
     2221 taaccaatta cggcatattc gcttagaaaa taatgacaac aaaccggtta ccaattcaag
     2281 ggacactcaa gaggtacccc tagaaaaagc taagcaagtg cttaaaataa ttgctacttt
     2341 caagcatacc acctcaatct ttgatgactt tgcacattat gaaaagcgtc aagaagagga
     2401 ggaagccatg cgtagggaga gaaatagaaa caaacaactc gaggactaca aagaccatga
     2461 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2521 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2581 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2641 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2701 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2761 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2821 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2881 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2941 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3001 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3061 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3121 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3181 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     3241 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3301 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3361 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3421 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3721 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3781 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3841 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3961 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4141 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4201 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4261 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4321 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4381 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4441 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4501 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4561 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4621 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4681 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4741 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4801 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4861 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4921 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4981 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5041 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     5101 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5161 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5221 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5341 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5401 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5461 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5521 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6001 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6241 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7561 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     7621 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7681 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     7741 tgaagaatct gcttagg
//
