LOCUS       Exported                6640 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encode YTHDF1 with N-terminal GST tag for E. coli expression.
ACCESSION   .
VERSION     .
KEYWORDS    pGEx-4T-1-YTHDF1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Wang X, Lu Z, Gomez A, Hon GC, Yue Y, Han D, Fu Y, Parisien M, Dai 
            Q, Jia G, Ren B, Pan T, He C
  TITLE     N6-methyladenosine-dependent regulation of messenger RNA stability.
  JOURNAL   Nature. 2014 Jan 2;505(7481):117-20. doi: 10.1038/nature12730. Epub 
            2013 Nov 27.
  PUBMED    24284625
REFERENCE   2  (bases 1 to 6640)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6640
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(56..74)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        142..246
                     /gene="bla"
                     /label=AmpR promoter
     CDS             247..1107
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(465..484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1278..1866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1767..1786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2020..2037
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2110..2187
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2188..3270
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2207..2226)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        3319..3349
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3357..3373
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3381..3397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3381..3397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3393..3920
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3413..3430)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3413..3429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3422..3444)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        4022..4050
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4058..4074
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     CDS             4097..4750
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4708..4730
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4757..4774
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6529..6551)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
       61 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      121 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      181 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      241 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      301 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      361 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      421 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      481 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      541 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      601 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      661 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      721 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      781 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      841 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      901 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      961 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1021 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1081 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1141 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1201 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1261 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1321 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1381 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1801 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1861 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1921 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1981 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2041 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2101 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     2161 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     2221 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     2281 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     2341 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     2401 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     2461 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     2521 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     2581 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     2641 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     2701 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     2761 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     2821 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     2881 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     2941 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     3001 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     3061 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     3121 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     3181 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3241 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     3301 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     3361 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
     3421 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3481 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3541 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
     3601 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
     3661 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg
     3721 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     3781 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta
     3841 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     3901 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     3961 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     4021 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
     4081 acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     4141 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     4201 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     4261 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     4321 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     4381 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     4441 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     4501 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     4561 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     4621 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     4681 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     4741 tcctccaaaa tcggatctgg ttccgcgtgg atccccggaa ttcatgtcgg ccaccagcgt
     4801 ggacacccag agaacaaaag gacaagataa taaagtacaa aatggttcgt tacatcagaa
     4861 ggatacagtt catgacaatg actttgagcc ctaccttact ggacagtcaa atcagagtaa
     4921 cagttacccc tcaatgagcg acccctacct gtccagctat tacccgccgt ccattggatt
     4981 tccttactcc ctcaatgagg ctccgtggtc tactgcaggg gaccctccga ttccatacct
     5041 caccacctac ggacagctca gtaacggaga ccatcatttt atgcacgatg ctgtttttgg
     5101 gcagcctggg ggcctgggga acaacatcta tcagcacagg ttcaattttt tccctgaaaa
     5161 ccctgcgttc tcagcatggg ggacaagtgg gtctcaaggt cagcagaccc agagctccgc
     5221 gtatgggagc agctacacct accccccgag ctccctgggt ggcacggtgg ttgatgggca
     5281 gccaggcttt cacagcgaca ccctcagcaa ggcccccggg atgaacagcc tggagcaggg
     5341 catggttggc ctgaagattg gggacgtcag ctcctccgcc gtcaagacgg tgggctctgt
     5401 cgtcagcagc gtggcactga ctggtgtcct ttctggcaac ggtgggacaa atgtgaacat
     5461 gccagtttca aagccgacct cgtgggctgc cattgccagc aagcctgcaa aaccacagcc
     5521 taaaatgaaa acaaagagcg ggcctgtcat ggggggtggg ctgccccctc cacccataaa
     5581 gcataacatg gacattggca cctgggataa caaggggcct gtgccgaagg ccccagtccc
     5641 ccagcaggca ccctctccac aggctgcccc acagccccag caggtggctc agcctctccc
     5701 agcacagccc ccagctttgg ctcaaccgca gtatcagagc cctcagcagc caccccagac
     5761 ccgctgggtt gccccacgca acagaaacgc ggcgtttggg cagagcggag gggctggcag
     5821 cgatagcaac tctcctggaa acgtccagcc taattctgcc cccagcgtcg aatcccaccc
     5881 cgtccttgaa aaactgaagg ctgctcacag ctacaacccg aaagagtttg agtggaatct
     5941 gaaaagcggg cgtgtgttca tcatcaagag ctactctgag gacgacatcc accgctccat
     6001 taagtactcc atctggtgta gcacagagca cggcaacaag cgcctggaca gcgccttccg
     6061 ctgcatgagc agcaaggggc ccgtctacct gctcttcagc gtcaatggga gtgggcattt
     6121 ttgtggggtg gccgagatga agtcccccgt ggactacggc accagtgccg gggtctggtc
     6181 tcaggacaag tggaagggga agtttgatgt ccagtggatt tttgttaagg atgtacccaa
     6241 taaccagctc cggcacatca ggctggagaa taacgacaac aaaccggtca caaactcccg
     6301 ggacacccag gaggtgccct tagaaaaagc caagcaagtg ctgaaaatta tcagttccta
     6361 caagcacaca acctccatct tcgacgactt tgctcactac gagaagcgcc aggaggagga
     6421 ggaggtggtg cgcaaggaac ggcagagtcg aaacaaacaa tgactcgagc ggccgcatcg
     6481 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6541 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6601 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
//
