pCMV-mA1-BE3

LOCUS       pCMV-mA1-BE3                8532 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses mA1-BE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-mA1-BE3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8532)
  AUTHORS   Wang X, Li J, Wang Y, Yang V, Wei J, Wu J, Wang R, Huang X, Chen J, 
            Yang L
  TITLE     Efficient base editing in methylated regions with a human 
            APOBEC3A-Cas9 fusion.
  JOURNAL   Nat Biotechnol.,  2018
REFERENCE   2  (bases 1 to 8532)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8532
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     762..781
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        762..780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1529..5629
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5642..5890
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5903..5923
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5932..5949
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5972..5989)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5978..6202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6273..6289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6273..6289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6286..6308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6297..6313
                     /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)."
     promoter        complement(6321..6351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6366..6387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6504..6521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6675..7263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6755..6774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7434..8294)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8057..8076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8295..8399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8478..8497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ggtttagtga accgtcagat ccgctagaga tccgcggccg ctaatacgac tcactatagg
      781 gagagccgcc accatgagtt ccgagacagg ccctgtagct gttgatccca ctctgaggag
      841 aagaattgag ccccacgagt ttgaagtctt ctttgacccc cgggagcttc ggaaagagac
      901 ctgtctgctg tatgagatca actggggtgg aaggcacagt gtctggcgac acacgagcca
      961 aaacaccagc aaccacgttg aagtcaactt cttagaaaaa tttactacag aaagatactt
     1021 tcgtccgaac accagatgct ccattacctg gttcctgtcc tggagtccct gcggggagtg
     1081 ctccagggcc attacagagt ttctgagccg acacccctat gtaactctgt ttatttacat
     1141 agcacggctt tatcaccaca cggatcagcg aaaccgccaa ggactcaggg accttattag
     1201 cagcggtgtg actatccaga tcatgacaga gcaagagtat tgttactgct ggaggaattt
     1261 cgtcaactac cccccttcaa acgaagctta ttggccaagg tacccccatc tgtgggtgaa
     1321 actgtatgta ctggagctct actgcatcat tttaggactt ccaccctgtt taaaaatttt
     1381 aagaagaaag caacctcaac tcacgttttt cacaattact cttcaaacct gccattacca
     1441 aaggatacca ccccatctcc tttgggctac agggttgaaa agcggcagcg agactcccgg
     1501 gacctcagag tccgccacac ccgaaagtga taaaaagtat tctattggtt tagccatcgg
     1561 cactaattcc gttggatggg ctgtcataac cgatgaatac aaagtacctt caaagaaatt
     1621 taaggtgttg gggaacacag accgtcattc gattaaaaag aatcttatcg gtgccctcct
     1681 attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa cgaaccgctc ggagaaggta
     1741 tacacgtcgc aagaaccgaa tatgttactt acaagaaatt tttagcaatg agatggccaa
     1801 agttgacgat tctttctttc accgtttgga agagtccttc cttgtcgaag aggacaagaa
     1861 acatgaacgg caccccatct ttggaaacat agtagatgag gtggcatatc atgaaaagta
     1921 cccaacgatt tatcacctca gaaaaaagct agttgactca actgataaag cggacctgag
     1981 gttaatctac ttggctcttg cccatatgat aaagttccgt gggcactttc tcattgaggg
     2041 tgatctaaat ccggacaact cggatgtcga caaactgttc atccagttag tacaaaccta
     2101 taatcagttg tttgaagaga accctataaa tgcaagtggc gtggatgcga aggctattct
     2161 tagcgcccgc ctctctaaat cccgacggct agaaaacctg atcgcacaat tacccggaga
     2221 gaagaaaaat gggttgttcg gtaaccttat agcgctctca ctaggcctga caccaaattt
     2281 taagtcgaac ttcgacttag ctgaagatgc caaattgcag cttagtaagg acacgtacga
     2341 tgacgatctc gacaatctac tggcacaaat tggagatcag tatgcggact tatttttggc
     2401 tgccaaaaac cttagcgatg caatcctcct atctgacata ctgagagtta atactgagat
     2461 taccaaggcg ccgttatccg cttcaatgat caaaaggtac gatgaacatc accaagactt
     2521 gacacttctc aaggccctag tccgtcagca actgcctgag aaatataagg aaatattctt
     2581 tgatcagtcg aaaaacgggt acgcaggtta tattgacggc ggagcgagtc aagaggaatt
     2641 ctacaagttt atcaaaccca tattagagaa gatggatggg acggaagagt tgcttgtaaa
     2701 actcaatcgc gaagatctac tgcgaaagca gcggactttc gacaacggta gcattccaca
     2761 tcaaatccac ttaggcgaat tgcatgctat acttagaagg caggaggatt tttatccgtt
     2821 cctcaaagac aatcgtgaaa agattgagaa aatcctaacc tttcgcatac cttactatgt
     2881 gggacccctg gcccgaggga actctcggtt cgcatggatg acaagaaagt ccgaagaaac
     2941 gattactcca tggaattttg aggaagttgt cgataaaggt gcgtcagctc aatcgttcat
     3001 cgagaggatg accaactttg acaagaattt accgaacgaa aaagtattgc ctaagcacag
     3061 tttactttac gagtatttca cagtgtacaa tgaactcacg aaagttaagt atgtcactga
     3121 gggcatgcgt aaacccgcct ttctaagcgg agaacagaag aaagcaatag tagatctgtt
     3181 attcaagacc aaccgcaaag tgacagttaa gcaattgaaa gaggactact ttaagaaaat
     3241 tgaatgcttc gattctgtcg agatctccgg ggtagaagat cgatttaatg cgtcacttgg
     3301 tacgtatcat gacctcctaa agataattaa agataaggac ttcctggata acgaagagaa
     3361 tgaagatatc ttagaagata tagtgttgac tcttaccctc tttgaagatc gggaaatgat
     3421 tgaggaaaga ctaaaaacat acgctcacct gttcgacgat aaggttatga aacagttaaa
     3481 gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa cttatcaacg ggataagaga
     3541 caagcaaagt ggtaaaacta ttctcgattt tctaaagagc gacggcttcg ccaataggaa
     3601 ctttatgcag ctgatccatg atgactcttt aaccttcaaa gaggatatac aaaaggcaca
     3661 ggtttccgga caaggggact cattgcacga acatattgcg aatcttgctg gttcgccagc
     3721 catcaaaaag ggcatactcc agacagtcaa agtagtggat gagctagtta aggtcatggg
     3781 acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc gaaaatcaaa cgactcagaa
     3841 ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa gagggtatta aagaactggg
     3901 cagccagatc ttaaaggagc atcctgtgga aaatacccaa ttgcagaacg agaaacttta
     3961 cctctattac ctacaaaatg gaagggacat gtatgttgat caggaactgg acataaaccg
     4021 tttatctgat tacgacgtcg atcacattgt accccaatcc tttttgaagg acgattcaat
     4081 cgacaataaa gtgcttacac gctcggataa gaaccgaggg aaaagtgaca atgttccaag
     4141 cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag ctcctaaatg cgaaactgat
     4201 aacgcaaaga aagttcgata acttaactaa agctgagagg ggtggcttgt ctgaacttga
     4261 caaggccgga tttattaaac gtcagctcgt ggaaacccgc caaatcacaa agcatgttgc
     4321 acagatacta gattcccgaa tgaatacgaa atacgacgag aacgataagc tgattcggga
     4381 agtcaaagta atcactttaa agtcaaaatt ggtgtcggac ttcagaaagg attttcaatt
     4441 ctataaagtt agggagataa ataactacca ccatgcgcac gacgcttatc ttaatgccgt
     4501 cgtagggacc gcactcatta agaaataccc gaagctagaa agtgagtttg tgtatggtga
     4561 ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc gaacaggaga taggcaaggc
     4621 tacagccaaa tacttctttt attctaacat tatgaatttc tttaagacgg aaatcactct
     4681 ggcaaacgga gagatacgca aacgaccttt aattgaaacc aatggggaga caggtgaaat
     4741 cgtatgggat aagggccggg acttcgcgac ggtgagaaaa gttttgtcca tgccccaagt
     4801 caacatagta aagaaaactg aggtgcagac cggagggttt tcaaaggaat cgattcttcc
     4861 aaaaaggaat agtgataagc tcatcgctcg taaaaaggac tgggacccga aaaagtacgg
     4921 tggcttcgat agccctacag ttgcctattc tgtcctagta gtggcaaaag ttgagaaggg
     4981 aaaatccaag aaactgaagt cagtcaaaga attattgggg ataacgatta tggagcgctc
     5041 gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa ggttacaagg aagtaaaaaa
     5101 ggatctcata attaaactac caaagtatag tctgtttgag ttagaaaatg gccgaaaacg
     5161 gatgttggct agcgccggag agcttcaaaa ggggaacgaa ctcgcactac cgtctaaata
     5221 cgtgaatttc ctgtatttag cgtcccatta cgagaagttg aaaggttcac ctgaagataa
     5281 cgaacagaag caactttttg ttgagcagca caaacattat ctcgacgaaa tcatagagca
     5341 aatttcggaa ttcagtaaga gagtcatcct agctgatgcc aatctggaca aagtattaag
     5401 cgcatacaac aagcacaggg ataaacccat acgtgagcag gcggaaaata ttatccattt
     5461 gtttactctt accaacctcg gcgctccagc cgcattcaag tattttgaca caacgataga
     5521 tcgcaaacga tacacttcta ccaaggaggt gctagacgcg acactgattc accaatccat
     5581 cacgggatta tatgaaactc ggatagattt gtcacagctt gggggtgact ctggtggttc
     5641 tactaatctg tcagatatta ttgaaaagga gaccggtaag caactggtta tccaggaatc
     5701 catcctcatg ctcccagagg aggtggaaga agtcattggg aacaagccgg aaagcgatat
     5761 actcgtgcac accgcctacg acgagagcac cgacgagaat gtcatgcttc tgactagcga
     5821 cgcccctgaa tacaagcctt gggctctggt catacaggat agcaacggtg agaacaagat
     5881 taagatgctc tctggtggtt ctcccaagaa gaagaggaaa gtctaaccgg tcatcatcac
     5941 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     6001 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6061 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     6121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     6181 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg
     6241 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6301 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt
     6361 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6421 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6481 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6541 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7441 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7501 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7561 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7621 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7681 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7741 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7801 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7861 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7921 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7981 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8041 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8101 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8161 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8221 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8281 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8401 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc
     8461 ctagggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8521 cctgcttgtg tg
//