LOCUS       sequence-414001-       12126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains cloning site for molecularly-tagged Cas9 gRNA constructs, 
            inducible Cas9, rtTA-T2A-puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12126)
  AUTHORS   Nuttle X, Burt ND, Currall B, Moyses-Oliveira M, Mohajeri K, Bhavsar
            R, Lucente D, Yadav R, Tai DJC, Gusella JF, Talkowski ME
  TITLE     Parallelized engineering of mutational models using piggyBac 
            transposon delivery of CRISPR libraries.
  JOURNAL   Cell Rep Methods. 2023 Dec 5:100672. doi: 
            10.1016/j.crmeth.2023.100672.
  PUBMED    38091988
REFERENCE   2  (bases 1 to 12126)
  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: "Cell Rep 
            Methods. 2023 Dec 5:100672. doi: 10.1016/j.crmeth.2023.100672."
COMMENT     Sequence Label: pPB9v2
FEATURES             Location/Qualifiers
     source          1..12126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..67
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        554..802
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     554..574
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     725..744
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        877..1255
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    885..903
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    921..939
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    957..975
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    993..1011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1029..1047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1065..1083
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1101..1119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1164..1188
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1282..1291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1294..1314
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1339..5439
                     /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             1339..5439
                     /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             5440..5487
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5542..5561)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5586..5634
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5865..6076
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             6089..6357
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6391..7134
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             7135..7188
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7189..7788
                     /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(7189..7208)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7685..7705
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7821..7942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7858..7877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7912..7931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8711..8745
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9236..9252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9236..9252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9249..9271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9260..9276
                     /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(9284..9290,9291..9308,9309..9314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9329..9350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9467..9484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9638..10226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9718..9737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10397..11188,11189..11257))
                     /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     11020..11039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11258..11362)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11388..11843)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11525..11546)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11737..11756
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11970..11992
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11984..12001
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11985..12001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg tgtaaaattg
       61 acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa
      121 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
      181 tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt
      241 atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
      301 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
      361 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
      421 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaggcctc
      481 cacggccaga ctagaggtac caacgcggcc tttttacggt tcctggcctt ttgctggcct
      541 tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      601 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
      661 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      721 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      781 cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct aattcgctag ctaggtcttg
      841 aaaggagtgg gaattggctc cggtgcccgt cagctcgagt ttactcccta tcagtgatag
      901 agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga ctttactccc
      961 tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag aacgtatgac
     1021 cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta tcagtgatag
     1081 agaacgtata tccagtttac tccctatcag tgatagagaa cgtataagct ttaggcgtgt
     1141 acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc ctggagcaat
     1201 tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaaggtct
     1261 agagctagcg aattcgaatt tgccaccatg gccccaaaga agaagcggaa ggtcggtatc
     1321 cacggagtcc cagcagccga caagaagtac tccattgggc tcgatatcgg cacaaacagc
     1381 gtcggctggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg
     1441 ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc
     1501 ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga
     1561 aagaatcgga tctgctacct gcaggagatc ttcagcaacg agatggccaa ggtggacgac
     1621 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     1681 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     1741 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     1801 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     1861 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     1921 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     1981 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     2041 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     2101 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     2161 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     2221 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     2281 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     2341 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     2401 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     2461 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     2521 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     2581 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     2641 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     2701 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     2761 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     2821 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     2881 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     2941 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     3001 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     3061 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     3121 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     3181 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     3241 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     3301 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     3361 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     3421 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     3481 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     3541 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     3601 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     3661 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     3721 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     3781 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     3841 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     3901 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     3961 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     4021 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     4081 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     4141 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     4201 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     4261 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     4321 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     4381 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     4441 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     4501 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     4561 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     4621 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     4681 agcgataagc tgatcgccag aaagaaggac tgggacccca agaaatacgg cggattcgat
     4741 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     4801 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa
     4861 aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc
     4921 attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
     4981 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
     5041 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
     5101 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa
     5161 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
     5221 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
     5281 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg
     5341 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
     5401 tatgaaacaa gaatcgacct ctctcagctc ggtggagaca aaaggccggc ggccacgaaa
     5461 aaggccggcc aggcaaaaaa gaaaaagtga aagggttcga tccctaccgg ttagtaatga
     5521 gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5581 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     5641 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5701 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5761 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcga aatcggatcc
     5821 agctgaatct aagtggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     5881 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     5941 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     6001 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     6061 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     6121 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     6181 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     6241 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     6301 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatctaa
     6361 gctgtgaccg gcgcctactc tagagccgcc atgtctagac tggacaagag caaagtcata
     6421 aactctgctc tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc
     6481 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     6541 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg
     6601 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc
     6661 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     6721 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     6781 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     6841 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     6901 aaacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     6961 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac
     7021 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     7081 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cggggagggc
     7141 agaggaagtc tactaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac
     7201 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     7261 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag
     7321 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     7381 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     7441 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     7501 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     7561 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     7621 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     7681 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     7741 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg
     7801 gattacaaaa tttgtgggta aacttgttta ttgcagctta taatggttac aaataaagca
     7861 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     7921 ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt caattagtct
     7981 atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta atatttaaac
     8041 agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg ttgcagggtt
     8101 tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt tagccccgga
     8161 attgactgga ttcctttttt agggccgcgg cccattggta tggctttttc cccgtatccc
     8221 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc
     8281 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg
     8341 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa
     8401 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat
     8461 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     8521 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     8581 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     8641 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     8701 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat
     8761 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg
     8821 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag
     8881 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg
     8941 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg
     9001 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc
     9061 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac
     9121 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag
     9181 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat
     9241 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     9301 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     9361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     9421 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     9481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     9541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     9601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     9661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     9721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     9781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     9841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     9901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     9961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    10021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    10081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    10141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    10201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    10261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    10321 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    10381 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    10441 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    10501 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    10561 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    10621 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    10681 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    10741 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    10801 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    10861 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    10921 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    10981 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    11041 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    11101 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    11161 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    11221 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    11281 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    11341 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
    11401 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
    11461 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
    11521 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
    11581 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
    11641 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
    11701 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
    11761 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
    11821 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
    11881 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
    11941 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
    12001 tgagcgcgcc tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc
    12061 aaatgtgttt tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca
    12121 gctaga
//
