LOCUS       Exported                7033 bp ds-DNA     circular SYN 01-FEB-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2017 from SnapGene 4.0.4
            http://www.snapgene.com
COMMENT     pET28c(+)
FEATURES             Location/Qualifiers
     source          1..7033
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      12..467
                     /label=M13 origin
     rep_origin      29..335
                     /label=F1 ori
     CDS             complement(563..1375)
                     /label=KanR
     rep_origin      1421..2103
                     /label=ColE1 origin
     primer_bind     4906..4929
                     /label=pET28c-F
     primer_bind     4984..5003
                     /label=T7
     misc_binding    5003..5025
                     /label=LacO
     misc_feature    5057..5063
                     /label=rbs
     CDS             5071..6870
                     /codon_start=1
                     /label=His-YTHDF2
                     /translation="MGSSHHHHHHSSGLVPRGSHMSASSLLEQRPKGQGNKVQNGSVHQ
                     KDGLNDDDFEPYLSPQARPNNAYTAMSDSYLPSYYSPSIGFSYSLGEAAWSTGGDTAMP
                     YLTSYGQLSNGEPHFLPDAMFGQPGALGSTPFLGQHGFNFFPSGIDFSAWGNNSSQGQS
                     TQSSGYSSNYAYAPSSLGGAMIDGQSAFANETLNKAPGMNTIDQGMAALKLGSTEVASN
                     VPKVVGSAVGSGSITSNIVASNSLPPATIAPPKPASWADIASKPAKQQPKLKTKNGIAG
                     SSLPPPPIKHNMDIGTWDNKGPVAKAPSQALVQNIGQPTQGSPQPVGQQANNSPPVAQA
                     SVGQQTQPLPPPPPQPAQLSVQQQAAQPTRWVAPRNRGSGFGHNGVDGNGVGQSQAGSG
                     STPSEPHPVLEKLRSINNYNPKDFDWNLKHGRVFIIKSYSEDDIHRSIKYNIWCSTEHG
                     NKRLDAAYRSMNGKGPVYLLFSVNGSGHFCGVAEMKSAVDYNTCAGVWSQDKWKGRFDV
                     RWIFVKDVPNSQLRHIRLENNENKPVTNSRDTQEVPLEKAKQVLKIIASYKHTTSIFDD
                     FSHYEKRQEEEESVKKERQGRGK"
     misc_feature    5071..5073
                     /label=Open reading frame
     misc_feature    5083..5100
                     /label=His Tag
     misc_feature    5110..5127
                     /label=Thrombin cleavage site
     primer_bind     5123..5155
                     /label=YTHDF2-NdeI
                     /note="YTHDF2-NdeI-F GCAGCCATATGTCGGCCAGCAGCCTCTTGGAGC"
     misc_feature    5131..6870
                     /label=Human YTHDF2
                     /label=Human YTHDF2 (between NdeI and XhoI)
     primer_bind     6176..6201
                     /label=YTHDF2-370-BamH-F
     primer_bind     6360..6389
                     /label=YTHdomainDF2-BamH-F
     misc_feature    6361..6762
                     /label=YTH domain in DF2
     primer_bind     complement(6730..6762)
                     /label=YTHdomainDF2-Xho-R
     primer_bind     complement(6844..6880)
                     /label=YTHDF2-Xho-R
     primer_bind     complement(6844..6867)
                     /label=df2_pET_FLAG_insert_R
     primer_bind     6865..6888
                     /label=df2_pET_FLAG_insert_F
     misc_feature    6947..6966
                     /label=T7 Terminator sequence
     primer_bind     complement(6961..6984)
                     /label=pET28c-R
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
     5101 agcagcggcc tggtgccgcg cggcagccat atgtcggcca gcagcctctt ggagcagaga
     5161 ccaaaaggtc aaggaaacaa agtacaaaat ggatctgtac atcaaaagga tggattaaac
     5221 gatgatgatt ttgaacctta cttgagtcca caggcaaggc ccaataatgc atatactgcc
     5281 atgtcagatt cctacttacc cagttactac agtccctcca ttggcttctc ctattctttg
     5341 ggtgaagctg cttggtctac ggggggtgac acagccatgc cctacttaac ttcttatgga
     5401 cagctgagca acggagagcc ccacttccta ccagatgcaa tgtttgggca accaggagcc
     5461 ctaggtagca ctccatttct tggtcagcat ggttttaatt tctttcccag tgggattgac
     5521 ttctcagcat ggggaaataa cagttctcag ggacagtcta ctcagagctc tggatatagt
     5581 agcaattatg cttatgcacc tagctcctta ggtggagcca tgattgatgg acagtcagct
     5641 tttgccaatg agaccctcaa taaggctcct ggcatgaata ctatagacca agggatggca
     5701 gcactgaagt tgggtagcac agaagttgca agcaatgttc caaaagttgt aggttctgct
     5761 gttggtagcg ggtccattac tagtaacatc gtggcttcca atagtttgcc tccagccacc
     5821 attgctcctc caaaaccagc atcttgggct gatattgcta gcaagcctgc aaaacagcaa
     5881 cctaaactga agaccaagaa tggcattgca gggtcaagtc ttccgccacc cccgataaag
     5941 cataacatgg atattggaac ttgggataac aagggtcccg ttgcaaaagc cccctcacag
     6001 gctttggttc agaatatagg tcagccaacc caggggtctc ctcagcctgt aggtcagcag
     6061 gctaacaata gcccaccagt ggctcaggca tcagtagggc aacagacaca gccattgcct
     6121 ccacctccac cacagcctgc ccagctttca gtccagcaac aggcagctca gccaacccgc
     6181 tgggtagcac ctcggaaccg tggcagtggg ttcggtcata atggggtgga tggtaatgga
     6241 gtaggacagt ctcaggctgg ttctggatct actccttcag aaccccaccc agtgttggag
     6301 aagcttcggt ccattaataa ctataacccc aaagattttg actggaatct gaaacatggc
     6361 cgggttttca tcattaagag ctactctgag gacgatattc accgttccat taagtataat
     6421 atttggtgca gcacagagca tggtaacaag agactggatg ctgcttatcg ttccatgaac
     6481 gggaaaggcc ccgtttactt acttttcagt gtcaacggca gtggacactt ctgtggcgtg
     6541 gcagaaatga aatctgctgt ggactacaac acatgtgcag gtgtgtggtc ccaggacaaa
     6601 tggaagggtc gttttgatgt caggtggatt tttgtgaagg acgttcccaa tagccaactg
     6661 cgacacattc gcctagagaa caacgagaat aaaccagtga ccaactctag ggacactcag
     6721 gaagtgcctc tggaaaaggc taagcaggtg ttgaaaatta tagccagcta caagcacacc
     6781 acttccattt ttgatgactt ctcacactat gagaaacgcc aagaggaaga agaaagtgtt
     6841 aaaaaggaac gtcaaggtcg tgggaaatag ctcgagcacc accaccacca ccactgagat
     6901 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     6961 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     7021 actatatccg gat
//
