LOCUS       sequence-413996-       11745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains cloning site for Cas12a gRNA cassette, inducible Cas12a, 
            rtTA-T2A-puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11745)
  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 11745)
  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: pPB12A
FEATURES             Location/Qualifiers
     source          1..11745
                     /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        547..925
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    555..573
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    591..609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    627..645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    663..681
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    699..717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    735..753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    771..789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     834..858
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             968..4888
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKYFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      4608..4617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4889..4936
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5161..5180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5205..5253
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5484..5695
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5708..5976
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6010..6753
                     /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             6754..6807
                     /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             6808..7407
                     /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(6808..6827)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7304..7324
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7440..7561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7477..7496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7531..7550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8330..8364
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8855..8871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8855..8871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8868..8890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8879..8895
                     /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(8903..8909,8910..8927,8928..8933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8948..8969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9086..9103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9257..9845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9337..9356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10016..10807,10808..10876))
                     /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     10639..10658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10877..10981)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11007..11462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11144..11165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11356..11375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11589..11611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11603..11620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11604..11620
                     /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 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
      541 cagctcgagt ttactcccta tcagtgatag agaacgtatg aagagtttac tccctatcag
      601 tgatagagaa cgtatgcaga ctttactccc tatcagtgat agagaacgta taaggagttt
      661 actccctatc agtgatagag aacgtatgac cagtttactc cctatcagtg atagagaacg
      721 tatctacagt ttactcccta tcagtgatag agaacgtata tccagtttac tccctatcag
      781 tgatagagaa cgtataagct ttaggcgtgt acggtgggcg cctataaaag cagagctcgt
      841 ttagtgaacc gtcagatcgc ctggagcaat tccacaacac ttttgtctta taccaacttt
      901 ccgtaccact tcctaccctc gtaaaggtct agagctagcg tttaaactta agcttggtac
      961 cgccaccatg acacagttcg agggctttac caacctgtat caggtgagca agacactgcg
     1021 gtttgagctg atcccacagg gcaagaccct gaagcacatc caggagcagg gcttcatcga
     1081 ggaggacaag gcccgcaatg atcactacaa ggagctgaag cccatcatcg atcggatcta
     1141 caagacctat gccgaccagt gcctgcagct ggtgcagctg gattgggaga acctgagcgc
     1201 cgccatcgac tcctatagaa aggagaaaac cgaggagaca aggaacgccc tgatcgagga
     1261 gcaggccaca tatcgcaatg ccatccacga ctacttcatc ggccggacag acaacctgac
     1321 cgatgccatc aataagagac acgccgagat ctacaagggc ctgttcaagg ccgagctgtt
     1381 taatggcaag gtgctgaagc agctgggcac cgtgaccaca accgagcacg agaacgccct
     1441 gctgcggagc ttcgacaagt ttacaaccta cttctccggc ttttatgaga acaggaagaa
     1501 cgtgttcagc gccgaggata tcagcacagc catcccacac cgcatcgtgc aggacaactt
     1561 ccccaagttt aaggagaatt gtcacatctt cacacgcctg atcaccgccg tgcccagcct
     1621 gcgggagcac tttgagaacg tgaagaaggc catcggcatc ttcgtgagca cctccatcga
     1681 ggaggtgttt tccttccctt tttataacca gctgctgaca cagacccaga tcgacctgta
     1741 taaccagctg ctgggaggaa tctctcggga ggcaggcacc gagaagatca agggcctgaa
     1801 cgaggtgctg aatctggcca tccagaagaa tgatgagaca gcccacatca tcgcctccct
     1861 gccacacaga ttcatccccc tgtttaagca gatcctgtcc gataggaaca ccctgtcttt
     1921 catcctggag gagtttaaga gcgacgagga agtgatccag tccttctgca agtacaagac
     1981 actgctgaga aacgagaacg tgctggagac agccgaggcc ctgtttaacg agctgaacag
     2041 catcgacctg acacacatct tcatcagcca caagaagctg gagacaatca gcagcgccct
     2101 gtgcgaccac tgggatacac tgaggaatgc cctgtatgag cggagaatct ccgagctgac
     2161 aggcaagatc accaagtctg ccaaggagaa ggtgcagcgc agcctgaagc acgaggatat
     2221 caacctgcag gagatcatct ctgccgcagg caaggagctg agcgaggcct tcaagcagaa
     2281 aaccagcgag atcctgtccc acgcacacgc cgccctggat cagccactgc ctacaaccct
     2341 gaagaagcag gaggagaagg agatcctgaa gtctcagctg gacagcctgc tgggcctgta
     2401 ccacctgctg gactggtttg ccgtggatga gtccaacgag gtggaccccg agttctctgc
     2461 ccggctgacc ggcatcaagc tggagatgga gccttctctg agcttctaca acaaggccag
     2521 aaattatgcc accaagaagc cctactccgt ggagaagttc aagctgaact ttcagatgcc
     2581 tacactggcc tctggctggg acgtgaataa ggagaagaac aatggcgcca tcctgtttgt
     2641 gaagaacggc ctgtactatc tgggcatcat gccaaagcag aagggcaggt ataaggccct
     2701 gagcttcgag cccacagaga aaaccagcga gggctttgat aagatgtact atgactactt
     2761 ccctgatgcc gccaagatga tcccaaagtg cagcacccag ctgaaggccg tgacagccca
     2821 ctttcagacc cacacaaccc ccatcctgct gtccaacaat ttcatcgagc ctctggagat
     2881 cacaaaggag atctacgacc tgaacaatcc tgagaaggag ccaaagaagt ttcagacagc
     2941 ctacgccaag aaaaccggcg accagaaggg ctacagagag gccctgtgca agtggatcga
     3001 cttcacaagg gattttctgt ccaagtatac caagacaacc tctatcgatc tgtctagcct
     3061 gcggccatcc tctcagtata aggacctggg cgagtactat gccgagctga atcccctgct
     3121 gtaccacatc agcttccaga gaatcgccga gaaggagatc atggatgccg tggagacagg
     3181 caagctgtac ctgttccaga tctataacaa gtactttgcc aagggccacc acggcaagcc
     3241 taatctgcac acactgtatt ggaccggcct gttttctcca gagaacctgg ccaagacaag
     3301 catcaagctg aatggccagg ccgagctgtt ctaccgccct aagtccagga tgaagaggat
     3361 ggcacaccgg ctgggagaga agatgctgaa caagaagctg aaggatcaga aaaccccaat
     3421 ccccgacacc ctgtaccagg agctgtacga ctatgtgaat cacagactgt cccacgacct
     3481 gtctgatgag gccagggccc tgctgcccaa cgtgatcacc aaggaggtgt ctcacgagat
     3541 catcaaggat aggcgcttta ccagcgacaa gttctttttc cacgtgccta tcacactgaa
     3601 ctatcaggcc gccaattccc catctaagtt caaccagagg gtgaatgcct acctgaagga
     3661 gcaccccgag acacctatca tcggcatcga tcggggcgag agaaacctga tctatatcac
     3721 agtgatcgac tccaccggca agatcctgga gcagcggagc ctgaacacca tccagcagtt
     3781 tgattaccag aagaagctgg acaacaggga gaaggagagg gtggcagcaa ggcaggcctg
     3841 gtctgtggtg ggcacaatca aggatctgaa gcagggctat ctgagccagg tcatccacga
     3901 gatcgtggac ctgatgatcc actaccaggc cgtggtggtg ctggagaacc tgaatttcgg
     3961 ctttaagagc aagaggaccg gcatcgccga gaaggccgtg taccagcagt tcgagaagat
     4021 gctgatcgat aagctgaatt gcctggtgct gaaggactat ccagcagaga aagtgggagg
     4081 cgtgctgaac ccataccagc tgacagacca gttcacctcc tttgccaaga tgggcaccca
     4141 gtctggcttc ctgttttacg tgcctgcccc atatacatct aagatcgatc ccctgaccgg
     4201 cttcgtggac cccttcgtgt ggaaaaccat caagaatcac gagagccgca agcacttcct
     4261 ggagggcttc gactttctgc actacgacgt gaaaaccggc gacttcatcc tgcactttaa
     4321 gatgaacaga aatctgtcct tccagagggg cctgcccggc tttatgcctg catgggatat
     4381 cgtgttcgag aagaacgaga cacagtttga cgccaagggc acccctttca tcgccggcaa
     4441 gagaatcgtg ccagtgatcg agaatcacag attcaccggc agataccggg acctgtatcc
     4501 tgccaacgag ctgatcgccc tgctggagga gaagggcatc gtgttcaggg atggctccaa
     4561 catcctgcca aagctgctgg agaatgacga ttctcacgcc atcgacacca tggtggccct
     4621 gatccgcagc gtgctgcaga tgcggaactc caatgccgcc acaggcgagg actatatcaa
     4681 cagccccgtg cgcgatctga atggcgtgtg cttcgactcc cggtttcaga acccagagtg
     4741 gcccatggac gccgatgcca atggcgccta ccacatcgcc ctgaagggcc agctgctgct
     4801 gaatcacctg aaggagagca aggatctgaa gctgcagaac ggcatctcca atcaggactg
     4861 gctggcctac atccaggagc tgcgcaacaa aaggccggcg gccacgaaaa aggccggcca
     4921 ggcaaaaaag aaaaagtaag aattcgattc gtcagtaggg ttgtaaaggt ttttcttttc
     4981 ctgagaaaac aaccttttgt tttctcaggt tttgcttttt ggcctttccc tagctttaaa
     5041 aaaaaaaagc aaaactcacc gaggcagttc cataggatgg caagatcctg gtattggtct
     5101 gcgaaatttg ggtcttcgag aagacctgat tagtaatgag tttaaacggg ggaggctaac
     5161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5221 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5281 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5341 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5401 cggcaggccc tgccatagca gatctgcgaa atcggatcca gctgaatcta agtggatctg
     5461 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     5521 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     5581 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     5641 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     5701 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     5761 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     5821 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     5881 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     5941 ctttgtttcg ttttctgttc tgcgccgtta cagatctaag ctgtgaccgg cgcctactct
     6001 agagccgcca tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc
     6061 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     6121 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     6181 gagatgctgg acaggcatca tacccactcc tgccccctgg aaggcgagtc atggcaagac
     6241 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
     6301 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     6361 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     6421 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     6481 agagagacac ctaccaccga ttctatgccc ccacttctga aacaagcaat tgagctgttc
     6541 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     6601 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
     6661 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     6721 cttgacgatt ttgaccttga catgctcccc ggggagggca gaggaagtct actaacatgc
     6781 ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt gcgcctcgcc
     6841 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6901 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6961 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     7021 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     7081 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     7141 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     7201 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     7261 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     7321 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     7381 tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg attacaaaat ttgtgggtaa
     7441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7561 atcatgtctg gaattgactc aaatgatgtc aattagtcta tcagaagctc atctggtctc
     7621 ccttccgggg gacaagacat ccctgtttaa tatttaaaca gcagtgttcc caaactgggt
     7681 tcttatatcc cttgctctgg tcaaccaggt tgcagggttt cctgtcctca caggaacgaa
     7741 gtccctaaag aaacagtggc agccaggttt agccccggaa ttgactggat tcctttttta
     7801 gggccgcggc ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     7861 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     7921 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     7981 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     8041 gggggggctg tccctgatat ctataacaag aaaatatata tataataagt tatcacgtaa
     8101 gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa
     8161 gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac
     8221 cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac
     8281 agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat
     8341 tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat cgtcgggtct
     8401 tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga ggaagaagcc
     8461 cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga tgactgctgc
     8521 tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg tgtggccatg
     8581 cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc gtcgcggctt
     8641 ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa caataccggc
     8701 gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc gctgggatat
     8761 tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg gataccccgt
     8821 gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8881 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8941 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     9001 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     9061 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     9121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     9181 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     9241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     9721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    10021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    10081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    10141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    10201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    10261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    10621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    10681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    10921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    10981 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
    11041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
    11101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
    11161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
    11221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
    11281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
    11341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
    11401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
    11461 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
    11521 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
    11581 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcct cgttcattca
    11641 cgtttttgaa cccgtggagg acgggcagac tcgcggtgca aatgtgtttt acagcgtgat
    11701 ggagcagatg aagatgctcg acacgctgca gaacacgcag ctaga
//
