LOCUS       V018434                 7115 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018434
VERSION     V018434
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             929..2605
                     /label="YTHDF1(NM_017798)"
                     /note="YTHDF1(NM_017798)"
                     /gene="YTHDF1"
     CDS             2612..2677
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2715..2939
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2985..3413
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3427..3756
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3823..4617
                     /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    4791..4912
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4961..4977)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4985..5001
                     /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(5009..5039)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5054..5075
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5363..5948)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6119..6979)
                     /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(6980..7084)
                     /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 gtttaaactt aagcttggta ccgccaccat gtcggccacc agcgtggaca cccagagaac
      961 aaaaggacaa gataataaag tacaaaatgg ttcgttacat cagaaggata cagttcatga
     1021 caatgacttt gagccctacc ttactggaca gtcaaatcag agtaacagtt acccctcaat
     1081 gagcgacccc tacctgtcca gctattaccc gccgtccatt ggatttcctt actccctcaa
     1141 tgaggctccg tggtctactg caggggaccc tccgattcca tacctcacca cctacggaca
     1201 gctcagtaac ggagaccatc attttatgca cgatgctgtt tttgggcagc ctgggggcct
     1261 ggggaacaac atctatcagc acaggttcaa ttttttccct gaaaaccctg cgttctcagc
     1321 atgggggaca agtgggtctc aaggtcagca gacccagagc tccgcgtatg ggagcagcta
     1381 cacctacccc ccgagctccc tgggtggcac ggtggttgat gggcagccag gctttcacag
     1441 cgacaccctc agcaaggccc ccgggatgaa cagcctggag cagggcatgg ttggcctgaa
     1501 gattggggac gtcagctcct ccgccgtcaa gacggtgggc tctgtcgtca gcagcgtggc
     1561 actgactggt gtcctttctg gcaacggtgg gacaaatgtg aacatgccag tttcaaagcc
     1621 gacctcgtgg gctgccattg ccagcaagcc tgcaaaacca cagcctaaaa tgaaaacaaa
     1681 gagcgggcct gtcatggggg gtgggctgcc ccctccaccc ataaagcata acatggacat
     1741 tggcacctgg gataacaagg ggcctgtgcc gaaggcccca gtcccccagc aggcaccctc
     1801 tccacaggct gccccacagc cccagcaggt ggctcagcct ctcccagcac agcccccagc
     1861 tttggctcaa ccgcagtatc agagccctca gcagccaccc cagacccgct gggttgcccc
     1921 acgcaacaga aacgcggcgt ttgggcagag cggaggggct ggcagcgata gcaactctcc
     1981 tggaaacgtc cagcctaatt ctgcccccag cgtcgaatcc caccccgtcc ttgaaaaact
     2041 gaaggctgct cacagctaca acccgaaaga gtttgagtgg aatctgaaaa gcgggcgtgt
     2101 gttcatcatc aagagctact ctgaggacga catccaccgc tccattaagt actccatctg
     2161 gtgtagcaca gagcacggca acaagcgcct ggacagcgcc ttccgctgca tgagcagcaa
     2221 ggggcccgtc tacctgctct tcagcgtcaa tgggagtggg catttttgtg gggtggccga
     2281 gatgaagtcc cccgtggact acggcaccag tgccggggtc tggtctcagg acaagtggaa
     2341 ggggaagttt gatgtccagt ggatttttgt taaggatgta cccaataacc agctccggca
     2401 catcaggctg gagaataacg acaacaaacc ggtcacaaac tcccgggaca cccaggaggt
     2461 gcccttagaa aaagccaagc aagtgctgaa aattatcagt tcctacaagc acacaacctc
     2521 catcttcgac gactttgctc actacgagaa gcgccaggag gaggaggagg tggtgcgcaa
     2581 ggaacggcag agtcgaaaca aacaactcga ggactacaaa gaccatgacg gtgattataa
     2641 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg
     2701 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2761 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2821 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2881 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2941 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3781 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4321 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4801 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4861 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4921 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4981 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5041 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5761 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5881 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6361 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7081 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
