LOCUS       sequence-201329-       10917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces lambda Red and Cpf1 for recombination and selection. The 
            plasmid is combined with a donor plasmid (with crRNA and template) 
            for rapid genome editing in E.coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10917)
  AUTHORS   Ao X, Yao Y, Li T, Yang TT, Dong X, Zheng ZT, Chen GQ, Wu Q, Guo Y
  TITLE     A Multiplex Genome Editing Method for Escherichia coli Based on 
            CRISPR-Cas12a.
  JOURNAL   Front Microbiol. 2018 Oct 9;9:2307. doi: 10.3389/fmicb.2018.02307. 
            eCollection 2018.
  PUBMED    30356638
REFERENCE   2  (bases 1 to 10917)
  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 
            Microbiol."; date: "2018-10-9"; volume: "9:2307. doi"; pages: " 
            10.3389/fmicb.2018.02307. eCollection 2018"
COMMENT     Sequence Label: p46Cpf1-OP2
FEATURES             Location/Qualifiers
     source          1..10917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(18..644)
                     /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     549..568
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        660..715
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    696..714
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             733..744
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             758..1174
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             1180..1965
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             1962..2642
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      2643..2887
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(3026..3976)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4024..4246
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      4767..4861
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4872..5750)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5777..6061
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5984..6003
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             6083..6094
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      10015..10086
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      10102..10129
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(10155..10814)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     10729..10748
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(10815..10917)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gtcgagcatg cgacgtctta agacccactt tcacatttaa gttgtttttc taatccgcat
       61 atgatcaatt caaggccgaa taagaaggct ggctctgcac cttggtgatc aaataattcg
      121 atagcttgtc gtaataatgg cggcatacta tcagtagtag gtgtttccct ttcttcttta
      181 gcgacttgat gctcttgatc ttccaatacg caacctaaag taaaatgccc cacagcgctg
      241 agtgcatata atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc taattgattt
      301 tcgagagttt catactgttt ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg
      361 cgatgactta gtaaagcaca tctaaaactt ttagcgttat tacgtaaaaa atcttgccag
      421 ctttcccctt ctaaagggca aaagtgagta tggtgcctat ctaacatctc aatggctaag
      481 gcgtcgagca aagcccgctt attttttaca tgccaataca atgtaggctg ctctacacct
      541 agcttctggg cgagtttacg ggttgttaaa ccttcgattc cgacctcatt aagcagctct
      601 aatgcgctgt taatcacttt acttttatct aatctagaca tcattaattc ctaatttttg
      661 ttgacactct atcgttgata gagttatttt accactccct atcagtgata gagaaaagaa
      721 ttcaaaagat ctaaagagga gaaaggctcg agaaaaaatg gatattaata ctgaaactga
      781 gatcaagcaa aagcattcac taaccccctt tcctgttttc ctaatcagcc cggcatttcg
      841 cgggcgatat tttcacagct atttcaggag ttcagccatg aacgcttatt acattcagga
      901 tcgtcttgag gctcagagct gggcgcgtca ctaccagcag ctcgcccgtg aagagaaaga
      961 ggcagaactg gcagacgaca tggaaaaagg cctgccccag cacctgtttg aatcgctatg
     1021 catcgatcat ttgcaacgcc acggggccag caaaaaatcc attacccgtg cgtttgatga
     1081 cgatgttgag tttcaggagc gcatggcaga acacatccgg tacatggttg aaaccattgc
     1141 tcaccaccag gttgatattg attcagaggt ataaaacgaa tgagtactgc actcgcaacg
     1201 ctggctggga agctggctga acgtgtcggc atggattctg tcgacccaca ggaactgatc
     1261 accactcttc gccagacggc atttaaaggt gatgccagcg atgcgcagtt catcgcatta
     1321 ctgatcgttg ccaaccagta cggccttaat ccgtggacga aagaaattta cgcctttcct
     1381 gataagcaga atggcatcgt tccggtggtg ggcgttgatg gctggtcccg catcatcaat
     1441 gaaaaccagc agtttgatgg catggacttt gagcaggaca atgaatcctg tacatgccgg
     1501 atttaccgca aggaccgtaa tcatccgatc tgcgttaccg aatggatgga tgaatgccgc
     1561 cgcgaaccat tcaaaactcg cgaaggcaga gaaatcacgg ggccgtggca gtcgcatccc
     1621 aaacggatgt tacgtcataa agccatgatt cagtgtgccc gtctggcctt cggatttgct
     1681 ggtatctatg acaaggatga agccgagcgc attgtcgaaa atactgcata cactgcagaa
     1741 cgtcagccgg aacgcgacat cactccggtt aacgatgaaa ccatgcagga gattaacact
     1801 ctgctgatcg ccctggataa aacatgggat gacgacttat tgccgctctg ttcccagata
     1861 tttcgccgcg acattcgtgc atcgtcagaa ctgacacagg ccgaagcagt aaaagctctt
     1921 ggattcctga aacagaaagc cgcagagcag aaggtggcag catgacaccg gacattatcc
     1981 tgcagcgtac cgggatcgat gtgagagctg tcgaacaggg ggatgatgcg tggcacaaat
     2041 tacggctcgg cgtcatcacc gcttcagaag ttcacaacgt gatagcaaaa ccccgctccg
     2101 gaaagaagtg gcctgacatg aaaatgtcct acttccacac cctgcttgct gaggtttgca
     2161 ccggtgtggc tccggaagtt aacgctaaag cactggcctg gggaaaacag tacgagaacg
     2221 acgccagaac cctgtttgaa ttcacttccg gcgtgaatgt tactgaatcc ccgatcatct
     2281 atcgcgacga aagtatgcgt accgcctgct ctcccgatgg tttatgcagt gacggcaacg
     2341 gccttgaact gaaatgcccg tttacctccc gggatttcat gaagttccgg ctcggtggtt
     2401 tcgaggccat aaagtcagct tacatggccc aggtgcagta cagcatgtgg gtgacgcgaa
     2461 aaaatgcctg gtactttgcc aactatgacc cgcgtatgaa gcgtgaaggc ctgcattatg
     2521 tcgtgattga gcgggatgaa aagtacatgg cgagttttga cgagatcgtg ccggagttca
     2581 tcgaaaaaat ggacgaggca ctggctgaaa ttggttttgt atttggggag caatggcgat
     2641 gacgcatcct cacgataata tccgggtagg cgcaatcact ttcgtctact ccgttacaaa
     2701 gcgaggctgg gtatttcccg gcctttctgt tatccgaaat ccactgaaag cacagcggct
     2761 ggctgaggag ataaataata aacgaggggc tgtatgcaca aagcatcttc tgttgagtta
     2821 agaacgagta tcgagatggc acatagcctt gctcaaattg gaatcaggtt tgtgccaata
     2881 ccagtagaaa cagacgaaga atccatggag ctcatatggt accggtatgg acagttttcc
     2941 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     3001 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     3061 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     3121 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     3181 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     3241 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     3301 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     3361 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     3421 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     3481 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     3541 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     3601 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     3661 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     3721 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     3781 cattttccct actgatgttc atcatctgaa cgtattggtt ataagtgaac gataccgtcc
     3841 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     3901 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     3961 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     4021 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
     4081 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     4141 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     4201 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     4261 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     4321 aaaaccctaa aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa
     4381 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     4441 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     4501 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     4561 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     4621 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     4681 gactggctaa tgcacccagt aaggcagcgg tatcatcaac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg cctagggact cctgttgata gatccagtaa tgacctcaga
     4801 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     4861 tcatcgattt attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     4921 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa gtagagttga
     4981 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     5041 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     5101 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     5161 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     5221 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     5281 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     5341 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     5401 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     5461 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     5521 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     5581 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     5641 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     5701 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     5761 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     5821 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     5881 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     5941 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     6001 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
     6061 tgtgcacgaa ttcaaaagat ctaaagagga gaaaggatct atgtctatct accaggaatt
     6121 cgttaacaaa tactctctgt ctaaaaccct gcgtttcgaa ctgatcccgc agggtaaaac
     6181 cctggaaaac atcaaagctc gtggtctgat cctggacgac gaaaaacgtg ctaaagacta
     6241 caaaaaagct aaacagatca tcgacaaata ccaccagttc ttcatcgaag aaatcctctc
     6301 gtctgtctgc atcagcgaag acctgctgca gaactactcg gacgtgtact tcaaactaaa
     6361 aaaatctgac gacgacaacc tgcagaaaga cttcaaatct gctaaagaca ccatcaaaaa
     6421 acagatctct gaatacatca aagactctga aaaattcaaa aacctgttca accagaacct
     6481 gatcgacgct aaaaaaggtc aggaatctga cctgatcctg tggctgaaac agtctaaaga
     6541 caacggtatc gaactgttca aagctaactc tgacatcacc gacatcgacg aagctctgga
     6601 aatcatcaaa tctttcaaag gttggaccac ctacttcaaa ggtttccacg aaaaccgtaa
     6661 aaacgtttac tcttctaacg acatcccgac ctctatcatc taccgtatcg ttgacgacaa
     6721 cctgccgaaa ttcctggaaa acaaagctaa atacgaatct ctgaaagaca aagctccgga
     6781 agctatcaac tacgaacaga tcaaaaaaga cctggctgaa gaactgacct tcgacatcga
     6841 ctacaaaacc tctgaagtta accagcgtgt tttctctctg gacgaagttt tcgaaatcgc
     6901 taacttcaac aactacctga accagtctgg tatcaccaaa ttcaacacca tcatcggtgg
     6961 taaattcgtt aacggtgaaa acaccaaacg taaaggtatc aacgaataca tcaacctgta
     7021 ctctcagcag atcaacgaca aaaccctgaa aaaatacaaa atgtctgttc tgttcaaaca
     7081 gatcctgtct gacaccgaat ctaaatcttt cgttatcgac aaactggaag acgactctga
     7141 cgttgttacc accatgcagt ctttctacga acagatcgct gctttcaaaa ccgttgaaga
     7201 aaaatctatc aaagaaaccc tgtctctgct gttcgacgac ctgaaagctc agaaactgga
     7261 cctgtctaaa atctacttca aaaacgacaa atctctgacc gacctctctc agcaggtatt
     7321 cgacgactac agcgttatcg gtaccgctgt tctggaatac atcacccagc agatcgctcc
     7381 gaaaaacctg gacaacccgt ctaaaaaaga acaggaactg atcgctaaaa aaaccgaaaa
     7441 agctaaatac ctgtctctgg aaaccatcaa actggctctg gaagaattca acaaacaccg
     7501 tgacatcgac aaacagtgcc gtttcgaaga aatcctggct aacttcgctg ctatcccgat
     7561 gatcttcgac gaaatcgctc agaacaaaga caacctggct cagatctcta tcaaatacca
     7621 gaaccagggt aaaaaagacc tgctgcaggc ttctgctgaa gacgacgtta aagctatcaa
     7681 agacctgctg gaccagacca acaacctgct gcacaaactg aaaatcttcc acatctctca
     7741 gtctgaagac aaagctaaca tcctggacaa agacgaacac ttctacctgg ttttcgaaga
     7801 atgctacttc gaactggcta acatcgttcc gctgtacaac aaaatccgta actacatcac
     7861 ccagaaaccg tactctgacg aaaaattcaa actgaacttc gaaaactcta ccctggctaa
     7921 cggttgggac aaaaacaaag aaccggacaa caccgctatc ctgttcatca aagacgacaa
     7981 atactacctg ggtgttatga acaaaaaaaa caacaaaatc ttcgacgaca aagctatcaa
     8041 agaaaacaaa ggtgaaggtt acaaaaaaat cgtttacaaa ctgctgccgg gtgctaacaa
     8101 aatgctgccg aaagttttct tctctgctaa atctatcaaa ttctacaacc cgtctgaaga
     8161 catcctgcgt atccgtaacc actctaccca caccaaaaac ggttctccgc agaaaggtta
     8221 cgaaaaattc gaattcaaca tcgaagactg ccgtaaattc atcgacttct acaaacagtc
     8281 tatctctaaa cacccggaat ggaaagactt cggtttccgt ttctctgaca cccagcgtta
     8341 caactctatc gacgaattct accgtgaagt tgaaaaccag ggttacaaac tgaccttcga
     8401 aaacatctct gaatcttaca tcgactctgt tgttaaccag ggtaaactgt acctgttcca
     8461 gatctacaac aaagacttct ctgcttactc taaaggtcgt ccgaacctgc acaccctgta
     8521 ctggaaagct ctgttcgacg aacgtaacct gcaggacgtt gtttacaaac tgaacggtga
     8581 agctgaactg ttctaccgta aacagtctat cccgaaaaaa atcacccacc cggctaaaga
     8641 agctatcgct aacaaaaaca aagacaaccc gaaaaaagaa tctgttttcg aatacgacct
     8701 gatcaaagac aaacgtttca ccgaagacaa attcttcttc cactgcccga tcaccatcaa
     8761 cttcaaatct tctggtgcta acaaattcaa cgacgaaatc aacctgctgc tgaaagaaaa
     8821 agctaacgac gttcacatcc tgtctatcga ccgtggtgaa cgtcacctgg cttactacac
     8881 cctggttgac ggtaaaggta acatcatcaa acaggacacc ttcaacatca tcggtaacga
     8941 ccgtatgaaa accaactacc acgacaaact ggctgctatc gaaaaagacc gtgactctgc
     9001 tcgtaaagac tggaaaaaaa tcaacaacat caaagaaatg aaagaaggtt acctgtctca
     9061 ggttgttcac gaaatcgcta aactggttat agaatacaat gcgatagttg tatttgaaga
     9121 cctgaacttc ggcttcaaac gtggtcgttt caaagttgaa aaacaggttt accagaaact
     9181 ggaaaaaatg ctgatcgaaa aactgaacta cctggttttc aaagacaacg aattcgacaa
     9241 aaccggtggt gttctgcgtg cttaccagct gaccgctccg ttcgaaacct tcaaaaaaat
     9301 gggtaaacag accggtatca tctactacgt tccggctggt ttcacctcta aaatctgccc
     9361 tgttactggc ttcgttaacc agctctaccc gaaatacgaa tctgtttcta aatctcagga
     9421 attcttctct aaattcgaca aaatctgcta caacctggac aaaggttact tcgaattctc
     9481 tttcgactac aaaaacttcg gtgacaaagc tgctaaaggt aaatggacca tcgcttcttt
     9541 cggttctcgt ctgatcaact tccgtaactc tgacaaaaac cacaactggg acacccgtga
     9601 agtttacccg accaaagaac tggaaaaact gctgaaagac tactctatcg aatacggtca
     9661 cggtgaatgc atcaaagctg ctatctgcgg tgaatctgac aaaaaattct tcgctaaact
     9721 gacctctgtt ctgaacacca tcctgcagat gcgtaactct aaaaccggta ccgaactgga
     9781 ctacctgatc tctccggttg ctgacgttaa cggtaacttc ttcgactctc gtcaggctcc
     9841 gaaaaacatg ccgcaggacg ctgacgctaa cggtgcttac cacatcggtc tgaaaggtct
     9901 gatgctgctg ggtcgtatca aaaacaacca ggaaggtaaa aaactgaacc tggttatcaa
     9961 aaacgaagaa tacttcgaat tcgttcagaa ccgtaacaac taataaccag gcatcaaata
    10021 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
    10081 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgg tgcacaagcg
    10141 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
    10201 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
    10261 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
    10321 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
    10381 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
    10441 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
    10501 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
    10561 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
    10621 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
    10681 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
    10741 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
    10801 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
    10861 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatca
//
