LOCUS       sequence-232947-       14822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial genome editing plasmid with kanamycin resistance marker 
            and using sacB as a counter-selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14822)
  AUTHORS   Niu TC, Lin GM, Xie LR, Wang ZQ, Xing WY, Zhang JY, Zhang CC
  TITLE     Expanding the potential of CRISPR-Cpf1 based genome editing 
            technology in the cyanobacterium Anabaena PCC 7120.
  JOURNAL   ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.
  PUBMED    30525474
REFERENCE   2  (bases 1 to 14822)
  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. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437."
COMMENT     Sequence Label: pCpf1b
FEATURES             Location/Qualifiers
     source          1..14822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      496..890
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            1232..1319
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2558..3529
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             4040..4879
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             4866..5717
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     promoter        5948..6393
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(6394..6480,6481..7815)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLEVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     8306..8325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        join(8429..8434,8435..8452,8453..8458)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        12382..12416
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(12484..12780)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     12630..12652
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12644..12661
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12645..12661
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12671..12690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12671..12689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12737..12757)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12737..12755)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12776..12792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12776..12792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12789..12811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12800..12816
                     /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(join(12824..12830,12831..12848,12849..12854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12869..12890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(13645..14460)
                     /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(13645..14460)
                     /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     14368..14387
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     14452..14471
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcattaaacg cgtattcagg ctgaccctgc gcgctgcgca gggctttatt gattccattt
       61 ttacactgat gaatgttccg ttgcgctgcc cggattacag ctgaaagcga ccaggtgctc
      121 ggcgtggcaa gactcgcagc gaacccgtag aaagccatgc tccagccgcc cgcattggag
      181 aaattcttca aattcccgtt gcacatagcc cggcaattcc tttccctgct ctgccataag
      241 cgcagcgaat gccgggtaat actcgtcaac gatctgatag agaagggttt gctcgggtcg
      301 gtggctctgg taacgaccag tatcccgatc ccggctggcc gtcctggccg ccacatgagg
      361 catgttccgc gtccttgcaa tactgtgttt acatacagtc tatcgcttag cggaaagttc
      421 ttttaccctc agccgaaatg cctgccgttg ctagacattg ccagccagtg cccgtcactc
      481 ccgtactaac tgtcacgaac ccctgcaata actgtcacgc ccccctgcaa taactgtcac
      541 gaacccctgc aataactgtc acgcccccaa acctgcaaac ccagcagggg cgggggctgg
      601 cggggtgttg gaaaaatcca tccatgatta tctaagaata atccactagg cgcggttatc
      661 agcgcccttg tggggcgctg ctgcccttgc ccaatatgcc cggccagagg ccggatagct
      721 ggtctattcg ctgcgctagg ctacacaccg ccccaccgct gcgcggcagg gggaaaggcg
      781 ggcaaagccc gctaaacccc acaccaaacc ccgcagaaat acgctggagc gcttttagcc
      841 gctttagcgg cctttccccc tacccgaagg gtgggggcgc gtgtgcagcc ccgcagggcc
      901 tgtctcggtc gatcattcag cccggctcat ccttctggcg tggcggcaga ccgaacaagg
      961 cgcggtcgtg gtcgcgttca aggtacgcat ccattgccgc catgagccga tcctccggcc
     1021 actcgctgct gttcaccttg gccaaaatca tggcccccac cagcaccttg cgccttgttt
     1081 cgttcttgcg ctcttgctgc tgttcccttg cccgcacccg ctgaatttcg gcattgattc
     1141 gcgctcgttg ttcttcgagc ttggccagcc gatccgccgc cttgttgctc cccttaacca
     1201 tcttgacacc ccattgttaa tgtgctgtct cgtaggctat catggaggca cagcggcggc
     1261 aatcccgacc ctactttgta ggggagggcg cacttaccgg tttctcttcg agaaactggc
     1321 ctaacggcca cccttcgggc ggtgcgctct ccgagggcca ttgcatggag ccgaaaagca
     1381 aaagcaacag cgaggcagca tggcgattta tcaccttacg gcgaaaaccg gcagcaggtc
     1441 gggcggccaa tcggccaggg ccaaggccga ctacatccag cgcgaaggca agtatgcccg
     1501 cgacatggat gaagtcttgc acgccgaatc cgggcacatg ccggagttcg tcgagcggcc
     1561 cgccgactac tgggatgctg ccgacctgta tgaacgcgcc aatgggcggc tgttcaagga
     1621 ggtcgaattt gccctgccgg tcgagctgac cctcgaccag cagaaggcgc tggcgtccga
     1681 gttcgcccag cacctgaccg gtgccgagcg cctgccgtat acgctggcca tccatgccgg
     1741 tggcggcgag aacccgcact gccacctgat gatctccgag cggatcaatg acggcatcga
     1801 gcggcccgcc gctcagtggt tcaagcggta caacggcaag accccggaga agggcggggc
     1861 acagaagacc gaagcgctca agcccaaggc atggcttgag cagacccgcg aggcatgggc
     1921 cgaccatgcc aaccgggcat tagagcgggc tggccacgac gcccgcattg accacagaac
     1981 acttgaggcg cagggcatcg agcgcctgcc cggtgttcac ctggggccga acgtggtgga
     2041 gatggaaggc cggggcatcc gcaccgaccg ggcagacgtg gccctgaaca tcgacaccgc
     2101 caacgcccag atcatcgact tacaggaata ccgggaggca atagaccatg aacgcaatcg
     2161 acagagtgaa gaaatccaga ggcatcaacg agttagcgga gcagatcgaa ccgctggccc
     2221 agagcatggc gacactggcc gacgaagccc ggcaggtcat gagccagacc cagcaggcca
     2281 gcgaggcgca ggcggcggag tggctgaaag cccagcgcca gacaggggcg gcatgggtgg
     2341 agctggccaa agagttgcgg gaggtagccg ccgaggtgag cagcgccgcg cagagcgccc
     2401 ggagcgcgtc gcgggggtgg cactggaagc tatggctaac cgtgatgctg gcttccatga
     2461 tgcctacggt ggtgctgctg atcgcatcgt tgctcttgct cgacctgacg ccactgacaa
     2521 ccgaggacgg ctcgatctgg ctgcgcttgg tggcccgatg aagaacgaca ggactttgca
     2581 ggccataggc cgacagctca aggccatggg ctgtgagcgc ttcgatatcg gcgtcaggga
     2641 cgccaccacc ggccagatga tgaaccggga atggtcagcc gccgaagtgc tccagaacac
     2701 gccatggctc aagcggatga atgcccaggg caatgacgtg tatatcaggc ccgccgagca
     2761 ggagcggcat ggtctggtgc tggtggacga cctcagcgag tttgacctgg atgacatgaa
     2821 agccgagggc cgggagcctg ccctggtagt ggaaaccagc ccgaagaact atcaggcatg
     2881 ggtcaaggtg gccgacgccg caggcggtga acttcggggg cagattgccc ggacgctggc
     2941 cagcgagtac gacgccgacc cggccagcgc cgacagccgc cactatggcc gcttggcggg
     3001 cttcaccaac cgcaaggaca agcacaccac ccgcgccggt tatcagccgt gggtgctgct
     3061 gcgtgaatcc aagggcaaga ccgccaccgc tggcccggcg ctggtgcagc aggctggcca
     3121 gcagatcgag caggcccagc ggcagcagga gaaggcccgc aggctggcca gcctcgaact
     3181 gcccgagcgg cagcttagcc gccaccggcg cacggcgctg gacgagtacc gcagcgagat
     3241 ggccgggctg gtcaagcgct tcggtgatga cctcagcaag tgcgacttta tcgccgcgca
     3301 gaagctggcc agccggggcc gcagtgccga ggaaatcggc aaggccatgg ccgaggccag
     3361 cccagcgctg gcagagcgca agcccggcca cgaagcggat tacatcgagc gcaccgtcag
     3421 caaggtcatg ggtctgccca gcgtccagct tgcgcgggcc gagctggcac gggcaccggc
     3481 accccgccag cgaggcatgg acaggggcgg gccagatttc agcatgtagt gcttgcgttg
     3541 gtactcacgc ctgttatact atgagtactc acgcacagaa gggggtttta tggaatacga
     3601 aaaaagcgct tcagggtcgg tctacctgat caaaagtgac aagggctatt ggttgcccgg
     3661 tggctttggt tatacgtcaa acaaggccga ggctggccgc ttttcagtcg ctgatatggc
     3721 cagccttaac cttgacggct gcaccttgtc cttgttccgc gaagacaagc ctttcggccc
     3781 cggcaagttt ctcggtgact gatatgaaag accaaaagga caagcagacc ggcgacctgc
     3841 tggccagccc tgacgctgta cgccaagcgc gatatgccga gcgcatgaag gccaaaggga
     3901 tgcgtcagcg caagttctgg ctgaccgacg acgaatacga ggcgctgcgc gagtgcctgg
     3961 aagaactcag agcggcgcag ggcgggggta gtgaccccgc cagcgcctaa ccaccaactg
     4021 cctgcaaagg aggcaatcaa tggctaccca taagcctatc aatattctgg aggcgttcgc
     4081 agcagcgccg ccaccgctgg actacgtttt gcccaacatg gtggccggta cggtcggggc
     4141 gctggtgtcg cccggtggtg ccggtaaatc catgctggcc ctgcaactgg ccgcacagat
     4201 tgcaggcggg ccggatctgc tggaggtggg cgaactgccc accggcccgg tgatctacct
     4261 gcccgccgaa gacccgccca ccgccattca tcaccgcctg cacgcccttg gggcgcacct
     4321 cagcgccgag gaacggcaag ccgtggctga cggcctgctg atccagccgc tgatcggcag
     4381 cctgcccaac atcatggccc cggagtggtt cgacggcctc aagcgcgccg ccgagggccg
     4441 ccgcctgatg gtgctggaca cgctgcgccg gttccacatc gaggaagaaa acgccagcgg
     4501 ccccatggcc caggtcatcg gtcgcatgga ggccatcgcc gccgataccg ggtgctctat
     4561 cgtgttcctg caccatgcca gcaagggcgc ggccatgatg ggcgcaggcg accagcagca
     4621 ggccagccgg ggcagctcgg tactggtcga taacatccgc tggcagtcct acctgtcgag
     4681 catgaccagc gccgaggccg aggaatgggg tgtggacgac gaccagcgcc ggttcttcgt
     4741 ccgcttcggt gtgagcaagg ccaactatgg cgcaccgttc gctgatcggt ggttcaggcg
     4801 gcatgacggc ggggtgctca agcccgccgt gctggagagg cagcgcaaga gcaagggggt
     4861 gccccgtggt gaagcctaag aacaagcaca gcctcagcca cgtccggcac gacccggcgc
     4921 actgtctggc ccccggcctg ttccgtgccc tcaagcgggg cgagcgcaag cgcagcaagc
     4981 tggacgtgac gtatgactac ggcgacggca agcggatcga gttcagcggc ccggagccgc
     5041 tgggcgctga tgatctgcgc atcctgcaag ggctggtggc catggctggg cctaatggcc
     5101 tagtgcttgg cccggaaccc aagaccgaag gcggacggca gctccggctg ttcctggaac
     5161 ccaagtggga ggccgtcacc gctgatgcca tggtggtcaa aggtagctat cgggcgctgg
     5221 caaaggaaat cggggcagag gtcgatagtg gtggggcgct caagcacata caggactgca
     5281 tcgagcgcct ttggaaggta tccatcatcg cccagaatgg ccgcaagcgg caggggtttc
     5341 ggctgctgtc ggagtacgcc agcgacgagg cggacgggcg cctgtacgtg gccctgaacc
     5401 ccttgatcgc gcaggccgtc atgggtggcg gccagcatgt gcgcatcagc atggacgagg
     5461 tgcgggcgct ggacagcgaa accgcccgcc tgctgcacca gcggctgtgt ggctggatcg
     5521 accccggcaa aaccggcaag gcttccatag ataccttgtg cggctatgtc tggccgtcag
     5581 aggccagtgg ttcgaccatg cgcaagcgcc gccagcgggt gcgcgaggcg ttgccggagc
     5641 tggtcgcgct gggctggacg gtaaccgagt tcgcggcggg caagtacgac atcacccggc
     5701 ccaaggcggc aggctgaccc cccccactct attgtaaaca agacattttt atcttttata
     5761 ttcaatggct tattttcctg ctaattggta ataccatgaa aaataccatg ctcagaaaag
     5821 gcttaacaat attttgaaaa attgcctact gagcgctgcc gcacagctcc ataggccgct
     5881 ttcctggctt tgcttccaga tgtatgctct tctgctctag ctagaggatc gatccttttt
     5941 aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt
     6001 tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca
     6061 gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt
     6121 ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct
     6181 aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc
     6241 taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt
     6301 tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg
     6361 ctaacacagt acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa
     6421 gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg
     6481 aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat
     6541 gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagttcc tgaattcgat
     6601 tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg aggtttggga cagctggcca
     6661 ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta
     6721 gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc
     6781 gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc
     6841 gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt
     6901 acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa
     6961 caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac
     7021 ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag
     7081 cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac
     7141 tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat
     7201 ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc
     7261 ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca
     7321 aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa
     7381 ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg
     7441 aacggcaaat ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt
     7501 acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag
     7561 ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt
     7621 acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat
     7681 atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccaag cttcctgctg
     7741 aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta
     7801 acagttaaca aataaaaacg caaaagaaaa tgccgatatc ctattggcat tttcttttat
     7861 ttcttatcaa cataaaggtg aatcccatat gaactatata aaagcaggca aatggctaac
     7921 cgtattccta accttttggt aatgactcca acttattgat agtgttttat gttcagataa
     7981 tgcccgatga ctttgtcatg cagcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8281 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     8341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8401 tccagttcga tgtaacccac tcgtgcactt tacactttat gcttccggct cgtatgttag
     8461 gaggtcttta tcatgtcaat ttatcaagaa tttgttaata aatatagttt aagtaaaact
     8521 ctaagatttg agttaatccc acagggtaaa acacttgaaa acataaaagc aagaggtttg
     8581 attttagatg atgagaaaag agctaaagac tacaaaaagg ctaaacaaat aattgataaa
     8641 tatcatcagt tttttataga ggagatatta agttcggttt gtattagcga agatttatta
     8701 caaaactatt ctgatgttta ttttaaactt aaaaagagtg atgatgataa tctacaaaaa
     8761 gattttaaaa gtgcaaaaga tacgataaag aaacaaatat ctgaatatat aaaggactca
     8821 gagaaattta agaatttgtt taatcaaaac cttatcgatg ctaaaaaagg gcaagagtca
     8881 gatttaattc tatggctaaa gcaatctaag gataatggta tagaactatt taaagccaat
     8941 agtgatatca cagatataga tgaggcgtta gaaataatca aatcttttaa aggttggaca
     9001 acttatttta agggttttca tgaaaataga aaaaatgttt atagtagcaa tgatattcct
     9061 acatctatta tttataggat agtagatgat aatttgccta aatttctaga aaataaagct
     9121 aagtatgaga gtttaaaaga caaagctcca gaagctataa actatgaaca aattaaaaaa
     9181 gatttggcag aagagctaac ctttgatatt gactacaaaa catctgaagt taatcaaaga
     9241 gttttttcac ttgatgaagt ttttgagata gcaaacttta ataattatct aaatcaaagt
     9301 ggtattacta aatttaatac tattattggt ggtaaatttg taaatggtga aaatacaaag
     9361 agaaaaggta taaatgaata tataaatcta tactcacagc aaataaatga taaaacactc
     9421 aaaaaatata aaatgagtgt tttatttaag caaattttaa gtgatacaga atctaaatct
     9481 tttgtaattg ataagttaga agatgatagt gatgtagtta caacgatgca aagtttttat
     9541 gagcaaatag cagcttttaa aacagtagaa gaaaaatcta ttaaagaaac actatcttta
     9601 ttatttgatg atttaaaagc tcaaaaactt gatttgagta aaatttattt taaaaatgat
     9661 aaatctctta ctgatctatc acaacaagtt tttgatgatt atagtgttat tggtacagcg
     9721 gtactagaat atataactca acaaatagca cctaaaaatc ttgataaccc tagtaagaaa
     9781 gagcaagaat taatagccaa aaaaactgaa aaagcaaaat acttatctct agaaactata
     9841 aagcttgcct tagaagaatt taataagcat agagatatag ataaacagtg taggtttgaa
     9901 gaaatacttg caaactttgc ggctattccg atgatatttg atgaaatagc tcaaaacaaa
     9961 gacaatttgg cacagatatc tatcaaatat caaaatcaag gtaaaaaaga cctacttcaa
    10021 gctagtgcgg aagatgatgt taaagctatc aaggatcttt tagatcaaac taataatctc
    10081 ttacataaac taaaaatatt tcatattagt cagtcagaag ataaggcaaa tattttagac
    10141 aaggatgagc atttttatct agtatttgag gagtgctact ttgagctagc gaatatagtg
    10201 cctctttata acaaaattag aaactatata actcaaaagc catatagtga tgagaaattt
    10261 aagctcaatt ttgagaactc gactttggct aatggttggg ataaaaataa agagcctgac
    10321 aatacggcaa ttttatttat caaagatgat aaatattatc tgggtgtgat gaataagaaa
    10381 aataacaaaa tatttgatga taaagctatc aaagaaaata aaggcgaggg ttataaaaaa
    10441 attgtttata aacttttacc tggcgcaaat aaaatgttac ctaaggtttt cttttctgct
    10501 aaatctataa aattttataa tcctagtgaa gatatactta gaataagaaa tcattccaca
    10561 catacaaaaa atggtagtcc tcaaaaagga tatgaaaaat ttgagtttaa tattgaagat
    10621 tgccgaaaat ttatagattt ttataaacag tctataagta agcatccgga gtggaaagat
    10681 tttggattta gattttctga tactcaaaga tataattcta tagatgaatt ttatagagaa
    10741 gttgaaaatc aaggctacaa actaactttt gaaaatatat cagagagcta tattgatagc
    10801 gtagttaatc agggtaaatt gtacctattc caaatctata ataaagattt ttcagcttat
    10861 agcaaagggc gaccaaatct acatacttta tattggaaag cgctgtttga tgagagaaat
    10921 cttcaagatg tggtttataa gctaaatggt gaggcagagc ttttttatcg taaacaatca
    10981 atacctaaaa aaatcactca cccagctaaa gaggcaatag ctaataaaaa caaagataat
    11041 cctaaaaaag agagtgtttt tgaatatgat ttaatcaaag ataaacgctt tactgaagat
    11101 aagtttttct ttcactgtcc tattacaatc aattttaaat ctagtggagc taataagttt
    11161 aatgatgaaa tcaatttatt gctaaaagaa aaagcaaatg atgttcatat attaagtata
    11221 gatagaggtg aaagacattt agcttactat actttggtag atggtaaagg caatatcatc
    11281 aaacaagata ctttcaacat cattggtaat gatagaatga aaacaaacta ccatgataag
    11341 cttgctgcaa tagagaaaga tagggattca gctaggaaag actggaaaaa gataaataac
    11401 atcaaagaga tgaaagaggg ctatctatct caggtagttc atgaaatagc taagctagtt
    11461 atagagtata atgctattgt ggtttttgag gatttaaatt ttggatttaa aagagggcgt
    11521 ttcaaggtag agaagcaggt ctatcaaaag ttagaaaaaa tgctaattga gaaactaaac
    11581 tatctagttt tcaaagataa tgagtttgat aaaactgggg gagtgcttag agcttatcag
    11641 ctaacagcac cttttgagac ttttaaaaag atgggtaaac aaacaggtat tatctactat
    11701 gtaccagctg gttttacttc aaaaatttgt cctgtaactg gttttgtaaa tcagttatat
    11761 cctaagtatg aaagtgtcag caaatctcaa gagttcttta gtaagtttga caagatttgt
    11821 tataaccttg ataagggcta ttttgagttt agttttgatt ataaaaactt tggtgacaag
    11881 gctgccaaag gcaagtggac tatagctagc tttgggagta gattgattaa ctttagaaat
    11941 tcagataaaa atcataattg ggatactcga gaagtttatc caactaaaga gttggagaaa
    12001 ttgctaaaag attattctat cgaatatggg catggcgaat gtatcaaagc agctatttgc
    12061 ggtgagagcg acaaaaagtt ttttgctaag ctaactagtg tcctaaatac tatcttacaa
    12121 atgcgtaact caaaaacagg tactgagtta gattatctaa tttcaccagt agcagatgta
    12181 aatggcaatt tctttgattc gcgacaggcg ccaaaaaata tgcctcaaga tgctgatgcc
    12241 aatggtgctt atcatattgg gctaaaaggt ctgatgctac taggtaggat caaaaataat
    12301 caagagggca aaaaactcaa tttggttatc aaaaatgaag agtattttga gttcgtgcag
    12361 aataggaata actaagaatt cttgacagct agctcagtcc taggtataat gctagcgctg
    12421 atttaggcaa aaacgggtct aagaacttta aataatttct actgttgtag atgagagcag
    12481 gtgtcagccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
    12541 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
    12601 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
    12661 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
    12721 cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
    12781 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    12841 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    12901 gctcactgcc cgctttccac ccacctgcaa aagtctaaga actttaaata atttctactg
    12961 ttgtagatgc tactattcct gtgccttcag ataattcagt ctaagaactt taaataattt
    13021 ctactgttgt agatgtctag agccttttgt attagtagcc ggtctaagaa ctttaaataa
    13081 tttctactgt tgtagattag cgatttatga aggtcatttt tttgtctagc tttaatgcgg
    13141 tagttggtac cgtcgatggc agaaattcga tatctagatc tcatggatcc gcaatggcaa
    13201 caacgttgcg ctcgactcta gaggatctgt aatccgggca gcgcaacgga acattcatca
    13261 gtgtaaaaat ggaatcaata aagccctgcg cagcgcgcag ggtcagcctg aatacgcgtt
    13321 taatgaccag cacagtcgtg atggcaaggt cagaatagcg ctgaggtctg cctcgtgaag
    13381 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
    13441 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
    13501 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
    13561 aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
    13621 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
    13681 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
    13741 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
    13801 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
    13861 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
    13921 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
    13981 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
    14041 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
    14101 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
    14161 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
    14221 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
    14281 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
    14341 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
    14401 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
    14461 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
    14521 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
    14581 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
    14641 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
    14701 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
    14761 aacaccttct tcacgaggca gacctcagcg ctattctgac cttgccatca cgactgtgct
    14821 gg
//
