LOCUS       sequence-157099-       11505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A modified version of plasmid pX330 (#42230): Added are a ccdB 
            stuffer and a Puromycin selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11505)
  AUTHORS   Pritchard CEJ, Kroese LJ, Huijbers IJ
  TITLE     Direct Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse 
            Zygotes.
  JOURNAL   Methods Mol Biol. 2017;1642:21-35. doi: 10.1007/978-1-4939-7169-5_2.
  PUBMED    28815491
REFERENCE   2  (bases 1 to 11505)
  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: "Methods Mol
            Biol."; date: "2017"; volume: "1642:21-35. doi"; pages: " 
            10.1007/978-1-4939-7169-5_2"
COMMENT     Sequence Label: pX330-Puro-ccdB
FEATURES             Location/Qualifiers
     source          1..11505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /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     complement(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1521..1540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1682..1930
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1682..1702
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1853..1872
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(1962..1967,1968..1985,1986..1992)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2046..2705
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2112..2131)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3047..3352
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3231..3250
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        3401..3476
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        3573..3858
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3860..4137
                     /label=chicken beta-actin promoter
     intron          4138..4366
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4378..4387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4387..4452
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4459..4479
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4504..8604
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4504..8604
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8605..8652
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8680..8697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8686..8893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        8911..9409
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8946..8969)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9342..9361
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9449..10048
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(9449..9468)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9945..9965
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     rep_origin      10664..11119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10751..10770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10961..10982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11136..11155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11255..11277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11315..11333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11401..11505
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     1741 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     1801 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     1861 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     1921 acgaaacacc gggtcttccg gccgcattag gcaccccagg ctttacactt tatgcttccg
     1981 gctcgtataa tgtgtggatt ttgagttagg atccggcgag attttcagga gctaaggaag
     2041 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2101 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2161 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2221 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2281 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2341 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2401 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2461 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2521 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2581 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct
     2641 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2701 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     2761 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggtg
     2821 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     2881 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     2941 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     3001 attgaaatga acggctcttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt
     3061 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3121 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3181 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     3241 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     3301 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     3361 ctccgttata cacagccagt ctgcaggtcg aggaagacct gttttagagc tagaaatagc
     3421 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     3481 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     3541 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     3601 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     3661 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     3721 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     3781 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     3841 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     3901 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     3961 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     4021 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     4081 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     4141 gtcgctgcga cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     4201 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     4261 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt
     4321 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc
     4381 accatggact ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac
     4441 gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca
     4501 gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
     4561 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     4621 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     4681 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     4741 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     4801 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     4861 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     4921 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     4981 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     5041 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     5101 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     5161 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     5221 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     5281 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     5341 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     5401 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     5461 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     5521 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     5581 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     5641 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     5701 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     5761 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     5821 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     5881 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     5941 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     6001 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     6061 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     6121 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     6181 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     6241 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     6301 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     6361 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     6421 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     6481 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     6541 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     6601 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     6661 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     6721 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     6781 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     6841 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     6901 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     6961 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     7021 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     7081 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     7141 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     7201 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     7261 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     7321 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     7381 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     7441 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     7501 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     7561 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     7621 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     7681 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     7741 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     7801 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     7861 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     7921 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     7981 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     8041 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     8101 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     8161 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     8221 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     8281 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     8341 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     8401 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     8461 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     8521 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     8581 gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca
     8641 aaaaagaaaa agtaagaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt
     8701 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     8761 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     8821 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     8881 ggcatgctgg ggagcggccg caattctacc gggtagggga ggcgcttttc ccaaggcagt
     8941 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     9001 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     9061 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     9121 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     9181 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     9241 tcgctttctg ggctcagcag ctgggaaggg tgggtccggg ggcgggctca ggggcgggct
     9301 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     9361 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcatcca
     9421 tctagatctc gagcagctga agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     9481 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     9541 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     9601 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     9661 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     9721 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     9781 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     9841 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     9901 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     9961 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
    10021 gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga
    10081 aaggagcgca cgaccccatg catcgatgat atcagatccc cgggatgcag aaattgatga
    10141 tctattaaac aataaagatg tccactaaaa tggaagtttt tcctgtcata ctttgttaag
    10201 aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg ggggtggggg
    10261 tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat tgctttatga
    10321 taatgtttca tagttggata tcataattta aacaagcaaa accaaattaa gggccagctc
    10381 attcctccca ctcatgatct atagatctat agatctctcg tgggatcatt gtttttctct
    10441 tgattcccac tttgtggttc taagtactgt ggtttccaaa tgtgtcagtt tcatagcctg
    10501 aagaacgaga tcagcagcct ctgttccaca tacacttcat tctcagtatt gttttgccaa
    10561 gttctaattc catcagaagc tcctgcaggg gcgcctgatg cggtattttc tccttacgca
    10621 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
    10681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    10741 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10861 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10981 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
    11041 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
    11101 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
    11161 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
    11221 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
    11281 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
    11341 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
    11401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    11461 caataaccct gataaatgct tcaataatat tgaaaaagga agagt
//
