pJC005.em

LOCUS      pJC005.em             11031 bp ds-DNA     circular SYN 29-MAR-2022
DEFINITION  CRISPR-Cas12a genome editing of E. faecium.
ACCESSION   .
VERSION     .
KEYWORDS    pJC005.em
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11031)
  AUTHORS   Chua MJ, Collins J
  TITLE     Rapid, Efficient, and Cost-Effective Gene Editing of Enterococcus 
            faecium with CRISPR-Cas12a.
  JOURNAL   Microbiol Spectr. 2022 Feb 2:e0242721. doi: 
            10.1128/spectrum.02427-21.
  PUBMED    35107356
REFERENCE   2  (bases 1 to 11031)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 29, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11031
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(4706..5294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4786..4805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5611..5627
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5611..5627
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(5694..6317)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     6225..6244
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    6427..6445
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             6510..10430
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     primer_bind     complement(10681..10698)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10681..10697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10690..10712)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aaaaagatat aatgggagat aagacggttc gtgttcgtgc tgacttgcac catatcataa
       61 aaatcgaaac agcaaagaat ggcggaaacg taaaagaagt tatggaaata agacttagaa
      121 gcaaacttaa gagtgtgttg atagtgcagt atcttaaaat tttgtataat aggaattgaa
      181 gttaaattag atgctaaaaa tttgtaatta agaaggagtg attacatgaa caaaaatata
      241 aaatattctc aaaacttttt aacgagtgaa aaagtactca accaaataat aaaacaattg
      301 aatttaaaag aaaccgatac cgtttacgaa attggaacag gtaaagggca tttaacgacg
      361 aaactggcta aaataagtaa acaggtaacg tctattgaat tagacagtca tctattcaac
      421 ttatcgtcag aaaaattaaa actgaatact cgtgtcactt taattcacca agatattcta
      481 cagtttcaat tccctaacaa acagaggtat aaaattgttg ggagtattcc ttaccattta
      541 agcacacaaa ttattaaaaa agtggttttt gaaagccatg cgtctgacat ctatctgatt
      601 gttgaagaag gattctacaa gcgtaccttg gatattcacc gaacactagg gttgctcttg
      661 cacactcaag tctcgattca gcaattgctt aagctgccag cggaatgctt tcatcctaaa
      721 ccaaaagtaa acagtgtctt aataaaactt acccgccata ccacagatgt tccagataaa
      781 tattggaagc tatatacgta ctttgtttca aaatgggtca atcgagaata tcgtcaactg
      841 tttactaaaa atcagtttca tcaagcaatg aaacacgcca aagtaaacaa tttaagtacc
      901 gttacttatg agcaagtatt gtctattttt aatagttatc tattatttaa cgggaggaaa
      961 taattctatg agtcgctttt gtaaatttgg aaagttacac gttactaaag ggaatgtaga
     1021 taaattatta ggtatactac tgacagcttc caaggagcta aagaggtccc tagcgcttag
     1081 aatcgcttta ggaaacacga tccagtccaa taatcgtcga taaaaacttt tgaaaaaggt
     1141 tggtgaaatt acctactttt ggaataatca caaatcacaa gtgattaatc acaaatcaca
     1201 agtgattaat cacttgttta ttaagatatt aaaagctata atttaaataa agcgtgaatt
     1261 ttattacaca aaaagagggg ggagaaactt ggaactagca tttagagaaa gcttaaaaaa
     1321 gatgagaggt accaaatcaa aagaaaaatt ctcccaagaa ttagaaatga gtagatcaaa
     1381 ttattcacga atagaatcag gaaaatcaga tccaaccata aaaacactag aacaaattgc
     1441 aaagttaact aactcaacgc tagtagtgga tttaatccca aatgagccaa cagaaccaga
     1501 accagaaaca gaatcagaac aagtaacatt ggatttagaa atggaagaag aaaaaagcaa
     1561 tgacttcgtg tgaataatgc gcgaaatcgt tgcttatttt ttttaaaagc ggtatactag
     1621 atataacgaa acaacgaact gaatagaaac gaaaaaagag ccatgacaca tttataaaat
     1681 gtttgacgac attttataaa tgcatagccc gataagattg ccaaaccaac gcttatcagt
     1741 tagtcagatg aactcttccc tcgtaagaag ttatttaatt aactttgttt gaagacggta
     1801 tataaccgta ctatcattat atagggaaat cagagagttt tcaagtatct aagctactga
     1861 atttaagaat tgttaagcaa tcaatcggaa atcgtttgat tgcttttttt gtattcattt
     1921 atagaaggtg gagtttgtat gaatcatgat gaatgtaaaa cttatataaa aaatagttta
     1981 ttggagataa gaaaattagc aaatatctat acactagaaa cgtttaagaa agagttagaa
     2041 aagagaaata tctacttaga aacaaaatca gataagtatt tttcttcgga gggggaagat
     2101 tatatatata agttaataga aaataacaaa ataatttatt cgattagtgg aaaaaaattg
     2161 acttataaag gaaaaaaatc tttttcaaaa catgcaatat tgaaacagtt gaatgaaaaa
     2221 gcaaaccaag ttaattaaac aacctatttt ataggattta taggaaagga gaacagctga
     2281 atgaatatcc cttttgttgt agaaactgtg cttcatgacg gcttgttaaa gtacaaattt
     2341 aaaaatagta aaattcgctc aatcactacc aagccaggta aaagcaaagg ggctattttt
     2401 gcgtatcgct caaaatcaag catgattggc ggtcgtggtg ttgttctgac ttccgaggaa
     2461 gcgattcaag aaaatcaaga tacatttaca cattggacac ccaacgttta tcgttatgga
     2521 acgtatgcag acgaaaaccg ttcatacacg aaaggacatt ctgaaaacaa tttaagacaa
     2581 atcaatacct tctttattga ttttgatatt cacacggcaa aagaaactat ttcagcaagc
     2641 gatattttaa caaccgctat tgatttaggt tttatgccta ctatgattat caaatctgat
     2701 aaaggttatc aagcatattt tgttttagaa acgccagtct atgtgacttc aaaatcagaa
     2761 tttaaatctg tcaaagcagc caaaataatt tcgcaaaata tccgagaata ttttggaaag
     2821 tctttgccag ttgatctaac gtgtaatcat tttggtattg ctcgcatacc aagaacggac
     2881 aatgtagaat tttttgatcc taattaccgt tattctttca aagaatggca agattggtct
     2941 ttcaaacaaa cagataataa gggctttact cgttcaagtc taacggtttt aagcggtaca
     3001 gaaggcaaaa aacaagtaga tgaaccctgg tttaatctct tattgcacga aacgaaattt
     3061 tcaggagaaa agggtttaat agggcgtaat aacgtcatgt ttaccctctc tttagcctac
     3121 tttagttcag gctattcaat cgaaacgtgc gaatataata tgtttgagtt taataatcga
     3181 ttagatcaac ccttagaaga aaaagaagta atcaaaattg ttagaagtgc ctattcagaa
     3241 aactatcaag gggctaatag ggaatacatt accattcttt gcaaagcttg ggtatcaagt
     3301 gatttaacca gtaaagattt atttgtccgt caagggtggt ttaaattcaa gaaaaaaaga
     3361 agcgaacgtc aacgtgttca tttgtcagaa tggaaagaag atttaatggc ttatattagc
     3421 gaaaaatcag atgtatacaa gccttattta gtgacgacca aaaaagagat tagagaagtg
     3481 ctaggcattc ctgaacggac attagataaa ttgctgaagg tactgaaggc gaatcaggaa
     3541 attttcttta agattaaacc aggaagaaat ggtggcattc aacttgctag tgttaaatca
     3601 ttgttgctat cgatcattaa agtaaaaaaa gaagaaaaag aaagctatat aaaggcgctg
     3661 acaaattctt ttgacttaga gcatacattc attcaagaga ctttaaacaa gctagcagaa
     3721 cgccctaaaa cggacacaca actcgatttg tttagctatg atacaggctg aaaataaaac
     3781 ccgcactatg ccattacatt tatatctatg atacgtgttt gttttttctt tgctgtttag
     3841 cgaatgatta gcagaaatat acagagtaag attttaatta attattaggg ggagaaggag
     3901 agagtagccc gaaaactttt agttggcttg gactgaacga agtgagggaa aggctactaa
     3961 aacgtcgagg ggcagtgaga gcgaagcgaa cacttgattt tttaattttc tatcttttat
     4021 aggtcattag agtatactta tttgtcctat aaactattta gcagcataat agatttattg
     4081 aataggtcat ttaagttgag catattagag gaggaaaatc ttggagaaat atttgaagaa
     4141 cccgattaca tggattggat tagttcttgt ggttacgtgg tttttaacta aaagtagtga
     4201 atttttgatt tttggtgtgt gtgtcttgtt gtcagtattt gctagtcaaa gtgattaaat
     4261 agaattctca tgtttgacag cttatcatcg gagctccgat gataagctgt caaacatgag
     4321 aattcccggg gatccttacg aaatcatcct gtggagctta gtaggtttag caagatggca
     4381 gcgcctaaat gtagaatgat aaaaggatta agagattaat ttccctaaaa atgataaaac
     4441 aagcgttttg aaagcgcttg tttttttggt ttgcagtcag agtagaatag aagtatcaaa
     4501 aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt
     4561 gctatgctgt taagaatggt tccaacgctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa agatgtgatc
     5461 cgtagcggta tgcgctccat caagaagagc gacttcgcgg agctggtgaa gtacatcacc
     5521 gacgagcaag gcaagaccga tcgggccccc tgcaggataa aaaaattgta gataaatttt
     5581 ataaaatagt tttatctaca atttttttat caggaaacag ctatgaccgc ggccgctgta
     5641 tccacataaa aataagaagc ctgcatttgc aggcttctta tttttatggt accttaagac
     5701 ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag gccgaataag
     5761 aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc
     5821 atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc
     5881 aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt
     5941 gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct
     6001 gtaggccgtg tacctaaatg tacttttgct ccatctcggt gacttagtaa agcacatcta
     6061 aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag
     6121 tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt
     6181 tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt
     6241 gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt
     6301 ttatctaatc tagacatcat taattcctcc tttttgttga cattatatca ttgatagagt
     6361 tatttgtcaa actagttttt tatttcgatg ccctggactt catgaaaaac taaaaaaaat
     6421 attgacactc tatcattgat agagtataat taaaataagc ttgatcgtag cgttaacaga
     6481 tctgagctcc tgcagtaaag gaggttttta tgacacagtt tgaaggcttc accaatctct
     6541 accaggtcag caagacgcta cgttttgagc ttatcccgca gggaaaaacc ctgaaacaca
     6601 ttcaggaaca ggggttcata gaggaagata aggcgcgtaa cgaccattat aaagaactga
     6661 agcctataat cgaccgtatt tataaaacgt acgcggatca gtgcctgcag ctggttcagc
     6721 tggattggga gaatctgtcc gcggctattg atagctatcg caaagagaag accgaggaaa
     6781 cccgtaacgc actgattgaa gagcaggcga cctatcggaa tgcgatccat gattacttca
     6841 tcggccgcac cgacaacctg accgatgcaa ttaacaaacg tcacgcagag atttacaaag
     6901 gtctgtttaa agcagagtta ttcaatggca aggttctgaa acagctgggt acggtcacca
     6961 ccaccgaaca cgaaaacgca ctgctgagga gctttgataa atttaccaca tatttcagcg
     7021 gtttctatga aaatcgtaag aatgtattta gcgccgaaga tatttccacc gcaattcctc
     7081 atcgtattgt gcaggataat tttccgaagt ttaaagaaaa ttgtcatatt tttacccgtc
     7141 tgatcaccgc ggtaccgagc ctgcgagagc attttgaaaa cgttaagaaa gccattggaa
     7201 tttttgtcag taccagcatt gaagaagtgt tttcgttccc gttctataac caactgctga
     7261 cccagaccca gattgatctg tacaatcagc tgctgggggg cataagccgc gaggcaggta
     7321 ccgaaaagat aaagggactc aatgaggtgc tgaatctggc aattcagaag aatgatgaaa
     7381 cggctcatat cattgctagc ctgccgcatc gtttcattcc cctgtttaag caaatcctga
     7441 gcgatcgcaa tacactgagc tttatcctcg aagagtttaa atcggacgaa gaagttatcc
     7501 agagcttttg caaatacaaa accctgctgc ggaacgaaaa tgtgctggag accgctgaag
     7561 cactgtttaa tgaactgaac tcgatcgacc tcacccatat ttttatatcc cacaaaaaac
     7621 tggaaaccat aagcagcgct ctgtgtgacc attgggatac cctgcgcaac gccctgtatg
     7681 aacggcgtat cagcgagctg accgggaaaa tcaccaaatc cgcaaaggaa aaagttcagc
     7741 gtagtctgaa acacgaggac atcaacctgc aagaaattat tagcgcagca ggtaaagagc
     7801 tgagcgaagc attcaaacag aaaaccagcg aaatcctgag ccatgcccat gctgcactgg
     7861 atcagccgct gccgaccacc ctgaaaaaac aggaggaaaa ggagattctg aaaagccaac
     7921 tggacagcct gctgggcctg tatcacctgc tggactggtt tgcagtcgat gagagcaacg
     7981 aggttgatcc tgagttctcc gctcgtctga ccggaatcaa gctggagatg gaaccgagtc
     8041 tgtcgtttta caataaagcg cgtaattacg cgaccaagaa accgtatagc gtggaaaaat
     8101 tcaaactgaa ctttcagatg ccgacccttg caagcggatg ggacgttaac aaagaaaaaa
     8161 acaatggggc aattctgttt gtgaaaaatg gcctctatta tctgggtatc atgccgaaac
     8221 agaaagggcg ctacaaagcc ctgtcatttg agccgaccga gaaaacctca gagggtttcg
     8281 acaagatgta ctacgattat ttcccggatg cggcaaaaat gatacccaaa tgtagcaccc
     8341 aactgaaggc agttacagcc cactttcaga cccataccac cccgatcctg ctgtcgaaca
     8401 attttataga gccgctggaa attaccaaag agatttatga tctgaataat ccggaaaagg
     8461 agcccaagaa atttcagacg gcgtatgcaa aaaagaccgg ggatcagaaa ggttatcgtg
     8521 aagcgctgtg caaatggatt gactttaccc gtgactttct gtcaaaatat accaaaacga
     8581 cgagcattga tctgagcagc ctacgtccga gcagccaata taaggatctg ggcgaatatt
     8641 acgccgaact gaatccgctg ctctaccata tttccttcca acgaatcgct gaaaaagaaa
     8701 taatggacgc cgttgaaacc ggcaaactgt atctgtttca aatctacaac aaagatttcg
     8761 ccaaaggcca tcacggtaag ccgaacctgc ataccctgta ttggaccggt ctgtttagcc
     8821 cggagaatct ggccaaaacc agcatcaagc tgaacggaca ggcagaactg ttttaccgcc
     8881 ccaaaagccg tatgaaaagg atggcacacc gcctgggcga aaaaatgctg aataagaaac
     8941 tcaaagatca gaaaacgccg ataccggata ccctttatca ggagctgtat gattatgtta
     9001 accaccggct gagccatgac ctgagcgacg aagcgcgtgc actgctgccg aacgtgatta
     9061 ccaaggaagt ctcgcatgaa attattaaag atcggcgctt caccagtgat aaatttttct
     9121 tccatgtacc gatcaccctg aattatcaag ccgcaaatag cccttccaaa tttaatcaac
     9181 gcgtgaatgc gtacctgaaa gagcatccgg agaccccaat tattggcata gaccgaggag
     9241 aacgcaatct catttatatc accgtcattg atagcaccgg taagatcctg gaacagcgta
     9301 gcctgaatac cattcagcag tttgactacc agaaaaagct ggacaacaga gaaaaggaac
     9361 gtgtagccgc ccggcaggct tggagtgtgg tgggtactat caaggatctg aagcaggggt
     9421 atctctccca agttatccat gaaattgtcg atctaatgat tcactatcaa gcagtagtgg
     9481 tactggaaaa tctgaatttc ggtttcaaaa gcaaacgtac agggatcgct gaaaaagccg
     9541 tttatcagca gttcgagaaa atgctgatag acaagctgaa ttgcctggtt ctgaaagatt
     9601 atccggcaga gaaggtgggc ggtgtgctga acccgtacca gctgactgat caatttacga
     9661 gctttgcaaa aatgggaacg cagagcggtt tcctgttcta tgttccggcg ccatatacca
     9721 gcaagataga cccgctgaca ggtttcgtag atccgtttgt ctggaaaacc attaaaaatc
     9781 atgaaagtcg caaacatttt ctggagggct ttgattttct gcactatgac gtgaaaaccg
     9841 gcgacttcat tctgcatttt aaaatgaacc gtaatctgtc ctttcagcgc ggcctgcctg
     9901 gctttatgcc ggcgtgggac attgtttttg aaaagaatga gacacagttt gatgccaaag
     9961 gtaccccctt tattgcgggg aaacgcattg tgcccgttat agaaaatcac cgcttcaccg
    10021 gacggtatag ggacttgtac ccggcaaatg aattgatagc gctgctggag gagaaaggta
    10081 ttgtctttcg ggatggatca aacatcctgc cgaagctgct ggagaacgat gacagccacg
    10141 caatagacac catggtagcg ctgatccgaa gcgtgctgca gatgcgtaac agtaatgcgg
    10201 ctacggggga agactacatt aatagcccgg tccgtgatct gaacggcgtt tgtttcgata
    10261 gcagatttca aaatccggag tggccgatgg atgccgatgc caatggagct taccatatcg
    10321 ctctcaaagg tcagctccta ctgaaccatt tgaaagaatc aaaagatctg aaactgcaga
    10381 acggcatctc gaatcaggac tggctggcct acattcaaga actgagaaac taatatgacc
    10441 atgattacga attcgagctc ggtacccggg gatcctataa gttttaataa aactttaaat
    10501 agaaaaaggc ttctctcatg agaagtcttt tttatttaaa ataaatataa aataaaatag
    10561 aggctataaa tagcctctat tttatgtgag aaatccctaa ataaaaagat gccagtgtgc
    10621 tggaattcgt catcgcatga gtcagcgatg actcgaggcc tgcagacatg caagcttggc
    10681 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
    10741 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
    10801 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tagcataaaa ataagaagcc
    10861 tgcatttgca ggcttcttat ttttatggcg cgccgttctg aatccttagc taatggttca
    10921 acaggtaact atgacgaaga tagcaccctg gagcaaccag gaatgaatta ctatcccttt
    10981 tatcaagaag cgcacaaaaa gaaaaacgaa atgatacacc aatcagtgca a
//