LOCUS       sequence-396325-        7740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of the APOBEC1-YTHmut fusion protein for 
            protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7740)
  AUTHORS   Zhu H, Yin X, Holley CL, Meyer KD
  TITLE     Improved Methods for Deamination-Based m(6)A Detection.
  JOURNAL   Front Cell Dev Biol. 2022 Apr 27;10:888279. doi: 
            10.3389/fcell.2022.888279. eCollection 2022.
  PUBMED    35573664
REFERENCE   2  (bases 1 to 7740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Cell 
            Dev Biol."; date: "2022-04-27"; volume: "10:888279. doi"; pages: " 
            10.3389/fcell.2022.888279. eCollection 2022"
COMMENT     Sequence Label: pET-APOBEC1-YTHmut
FEATURES             Location/Qualifiers
     source          1..7740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     205..224
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        205..223
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    224..248
                     /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)."
     RBS             263..285
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             304..321
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             331..1431
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1405..1428
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1444..1464
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1468..2154
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             2716..2742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2808..2825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2878..2896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2892..2939
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2976..3431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3273..3294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3524..4339)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3524..4339)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4247..4266
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4331..4350
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4461..5049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4950..4969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5203..5220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5235..5374
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5300..5319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5460..5482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5476..5667)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6476..7558)
                     /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     7520..7539
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7559..7636)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
       61 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
      121 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
      181 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
      241 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catgggttct
      301 tctcaccatc accatcacca tggttcttct atgaaaatcg aagaaggtaa actggtaatc
      361 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
      421 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag
      481 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
      541 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
      601 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
      661 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg
      721 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
      781 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
      841 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
      901 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat
      961 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
     1021 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
     1081 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
     1141 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     1201 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     1261 gagttggcga aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc
     1321 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     1381 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taatgggatc
     1441 gaggaaaacc tgtacttcca atccaatatg agcagcgaaa ccggtccggt ggcggttgac
     1501 ccgaccctgc gtcgtcgtat tgagccgcac gagttcgaag tgttctttga tccgcgtgag
     1561 ctgcgtaagg aaacctgcct gctgtacgaa attaactggg gtggccgtca cagcatctgg
     1621 cgtcacacca gccagaacac caacaagcac gttgaggtga acttcatcga aaaatttacc
     1681 accgagcgtt acttctgccc gaacacccgt tgcagcatta cctggtttct gagctggagc
     1741 ccgtgcggtg aatgcagccg tgcgatcacc gagttcctga gccgttatcc gcacgttacc
     1801 ctgtttatct acattgcgcg tctgtatcac cacgcggacc cgcgtaaccg tcaaggtctg
     1861 cgtgatctga tcagcagcgg cgtgaccatc cagattatga ccgagcaaga aagcggttac
     1921 tgctggcgta acttcgttaa ctatagcccg agcaacgaag cgcattggcc gcgttacccg
     1981 cacctgtggg tgcgtctgta cgttctggag ctgtattgca tcattctggg cctgccgccg
     2041 tgcctgaaca ttctgcgtcg taagcagccg caactgacct tctttaccat cgcgctgcag
     2101 agctgccact accaacgtct gccgccgcac attctgtggg cgaccggtct gaagagcggc
     2161 agcgaaaccc cgggtaccag cgaaagcgcg accccggagg gtcgtgtttt tatcattaag
     2221 agctacagcg aagacgatat ccaccgtagc attaaatata acatctggtg cagcaccgag
     2281 cacggcaaca agcgtctgga cgcggcgtac cgtagcatga acggtaaagg cccggtgtat
     2341 ctgctgttca gcgttaacgg tagcggccac ttttgcggtg tggcggaaat gaaaagcgcg
     2401 gttgattaca acacctgcgc gggtgtgtgg agccaggaca agtggaaagg ccgtttcgat
     2461 gttcgttgga tttttgtgaa ggacgttccg aacagccaac tgcgtcacat ccgtctggag
     2521 aacaacgaaa acaaaccggt gaccaacagc cgtgataccc aggaagtgcc gctggaaaag
     2581 gcgaaacaag ttctgaagat cattgcgagc tacaaacaca ccaccagcat cttcgacgat
     2641 tttagccact atgagaagcg tcaggaagag gaagagagcg tgaagaagga gcgtcaaggt
     2701 cgtggcaaac tggagtaccc gtatgacgtt ccggattatg cgtaaattgg aagtggataa
     2761 cggatccgaa ttcgagcgcc gtcgacaagc ttgcggccgc actcgagcac caccaccacc
     2821 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2881 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2941 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     3001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3181 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3301 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aactagtaac
     3421 gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3481 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     3541 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3601 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     3661 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3721 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3781 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3841 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3901 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3961 acactgccag cgcatcaaca atgttttcac ctgaatcagg atattcttct aatacctgga
     4021 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4081 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4141 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     4201 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4261 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     4321 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     4381 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4441 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4561 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4621 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4681 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4741 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4801 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4861 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4921 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4981 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5041 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5101 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5161 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5221 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5281 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     5341 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5401 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5461 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     5521 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     5581 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     5641 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     5701 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     5761 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     5821 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     5881 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     5941 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     6001 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     6061 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     6121 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     6181 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     6241 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     6301 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     6361 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     6421 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     6481 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6541 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     6601 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     6661 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     6721 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     6781 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6841 tgggaacgat accctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6901 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6961 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     7021 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     7081 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     7141 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     7201 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     7261 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     7321 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     7381 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     7441 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     7501 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     7561 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     7621 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     7681 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
//
