LOCUS       sequence-181339-        6770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified from pCas9-CR4 (Addgene: 62655) to use the high-fidelity 
            version of Cas9, SpCas9-HF1 (N497A/R661A/Q695A/Q926A) from 
            Kleinstiver et al 2016..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6770)
  AUTHORS   Moreb EA, Hoover B, Yaseen A, Valyasevi N, Roecker Z, Menacho-Melgar
            R, Lynch MD
  TITLE     Managing the SOS Response for Enhanced CRISPR-Cas-Based 
            Recombineering in E. coli through Transient Inhibition of Host RecA 
            Activity.
  JOURNAL   ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.
  PUBMED    28915012
REFERENCE   2  (bases 1 to 6770)
  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: "ACS Synth 
            Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174."
COMMENT     Sequence Label: pEM-Cas9HF1
FEATURES             Location/Qualifiers
     source          1..6770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(14..640)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     545..564
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(653..687)
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     promoter        688..743
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    724..742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(779..2266,2267..2269,2270..2758,2759..2761,2762..2860,
                     2861..2863,2864..3553,3554..3556,3557..4882)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF1
                     /note="carries the mutations N497A, R661A, Q695A, and 
                     Q926A"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             779..4882
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4883..4915
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     terminator      4942..5013
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5029..5056
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5084..5101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5218..5763)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5298..5317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5877..5971)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5992..6651)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6566..6585
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6652..6754)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 agatatcgac gtcttaagac ccactttcac atttaagttg tttttctaat ccgcatatga
       61 tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag
      121 cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga
      181 cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg
      241 catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga
      301 gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat
      361 gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt
      421 ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt
      481 cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct
      541 tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg
      601 cgctgttaat cactttactt ttatctaatc tagacatcat taattcctaa ttgctagcat
      661 tgtacctagg actgagctag ccataaagtt gacactctat cgttgataga gttattttac
      721 cactccctat cagtgataga gaaaagaatt caaaagatct aaagaggact tcggatctat
      781 ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
      841 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
      901 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
      961 gactcgcctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta
     1021 tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact
     1081 tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa
     1141 tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa
     1201 attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat
     1261 gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt
     1321 ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat
     1381 taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg
     1441 attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct
     1501 cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga
     1561 tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca
     1621 aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt
     1681 actttcagat atcctaagag taaatactga aataactaag gctcccctat cagcttcaat
     1741 gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca
     1801 acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg
     1861 ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga
     1921 aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa
     1981 gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc
     2041 tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga
     2101 aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg
     2161 ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt
     2221 tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacagcgt ttgataaaaa
     2281 tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta
     2341 taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc
     2401 aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt
     2461 taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc
     2521 aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat
     2581 taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt
     2641 aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca
     2701 cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggagc
     2761 gttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga
     2821 ttttttgaaa tcagatggtt ttgccaatcg caattttatg gcgctgatcc atgatgatag
     2881 tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca
     2941 tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt
     3001 aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat
     3061 tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat
     3121 gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt
     3181 tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga
     3241 catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat
     3301 tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga
     3361 taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa
     3421 ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac
     3481 gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt
     3541 ggttgaaact cgcgcgatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac
     3601 taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa
     3661 attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta
     3721 ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata
     3781 tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat
     3841 gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa
     3901 tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc
     3961 tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc
     4021 cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca
     4081 gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc
     4141 tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta
     4201 ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa
     4261 agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt
     4321 tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata
     4381 tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca
     4441 aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca
     4501 ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca
     4561 gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat
     4621 tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc
     4681 aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc
     4741 cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga
     4801 agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga
     4861 tttgagtcag ctaggaggtg acgctgctaa cgacgaaaac tacgctctgg ctgcttaact
     4921 cgagtaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     4981 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     5041 ggtgggcctt tctgcgttta tacctaggga tatattccgc ttcctcgctc actgactcgc
     5101 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     5161 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     5221 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     5281 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     5341 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     5401 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     5461 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5521 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     5581 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     5641 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     5701 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     5761 caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct
     5821 tcaaatgtag cacctgaagt cagccccata cgatataagt tgttactagt gcttggattc
     5881 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     5941 gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc
     6001 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     6061 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     6121 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     6181 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     6241 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     6301 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     6361 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     6421 cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     6481 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     6541 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     6601 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     6661 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     6721 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
//
