LOCUS       sequence-413995-       11929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Cas3 gRNA cloning site, inducible Cas11-P2A-Cas6-T2A-Cas3, 
            rtTA-T2A-blast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11929)
  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 11929)
  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: pPB1163
FEATURES             Location/Qualifiers
     source          1..11929
                     /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        933..1311
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    941..959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    977..995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1013..1031
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1049..1067
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1085..1103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1121..1139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1157..1175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1220..1244
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1338..1347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1350..1370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1896..1916
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1926..1982
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1986..2006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2649..2669
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2679..2732
                     /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             2736..2756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5466..5486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5541..5560)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5585..5633
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5864..6075
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             6088..6356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6390..7133
                     /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             7134..7187
                     /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             7188..7586
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    7625..7746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7662..7681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7716..7735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8514..8548
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9039..9055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9039..9055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9052..9074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9063..9079
                     /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(9087..9093,9094..9111,9112..9117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9132..9153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9270..9287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9441..10029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9521..9540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10200..10991,10992..11060))
                     /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     10823..10842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11061..11165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11191..11646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11328..11349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11540..11559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11773..11795
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11787..11804
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11788..11804
                     /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 ccgtggatgt gttgtttgtg tgatactata aagttggtag
      841 attgtgactg gcttaacgag acgtttaccc gtctctaatt cgctagctag gtcttgaaag
      901 gagtgggaat tggctccggt gcccgtcagc tcgagtttac tccctatcag tgatagagaa
      961 cgtatgaaga gtttactccc tatcagtgat agagaacgta tgcagacttt actccctatc
     1021 agtgatagag aacgtataag gagtttactc cctatcagtg atagagaacg tatgaccagt
     1081 ttactcccta tcagtgatag agaacgtatc tacagtttac tccctatcag tgatagagaa
     1141 cgtatatcca gtttactccc tatcagtgat agagaacgta taagctttag gcgtgtacgg
     1201 tgggcgccta taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca
     1261 caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa aggtctagag
     1321 ctagcgaatt cgaatttgcc accatggccc caaagaagaa gagaaaggtg agatctgaag
     1381 ccgatgagat cgacgcaatg gcactgtaca gggcatggca gcagctggac aacggatctt
     1441 gcgcacagat caggcgcgtg agcgagcctg atgagctgag ggacatccca gccttctatc
     1501 ggctggtgca gccctttggc tgggagaatc ctagacacca gcaggccctg ctgaggatgg
     1561 tgttttgtct gagcgccggc aagaacgtga tccggcacca ggacaagaag agcgagcaga
     1621 ccacaggaat ctccctggga cgcgccctgg ccaattccgg ccggatcaac gagcggagaa
     1681 tcttccagct gatcagggcc gatcgcacag ccgacatggt gcagctgagg cgcctgctga
     1741 cccacgcaga gcctgtgctg gattggccac tgatggcccg catgctgaca tggtggggca
     1801 agcgggagag acagcagctg ctggaggact tcgtgctgac cacaaataag aacgccaagc
     1861 ggactgctga tggcagtgaa tttgagtcca ctagtccaaa gaagaagaga aaggtgggaa
     1921 gcggagcaac caacttcagc ctgctgaagc aggccggcga tgtggaggag aatccaggcc
     1981 ccatgccaaa gaagaagaga aaggtgacgc gtgaatacct gagcaaagtg atcatcgcaa
     2041 gggcatggtc cagggacctg tatcagctgc accagggcct gtggcacctg ttccctaata
     2101 gaccagatgc cgccagggac ttcctgtttc acgtggagaa gaggaacaca cccgagggct
     2161 gtcacgtgct gctgcagtcc gcccagatgc ccgtgagcac cgcagtggcc acagtgatca
     2221 agaccaagca ggtggagttc cagctgcaag tgggcgtgcc actgtacttt aggctgcgcg
     2281 ccaatcccat caagaccatc ctggataacc agaagcgcct ggactctaag ggcaatatca
     2341 agcggtgcag agtgcctctg atcaaggagg ccgagcagat cgcctggctg cagagaaagc
     2401 tgggcaacgc cgccagggtg gaggatgtgc accctatcag cgagcggcca cagtatttca
     2461 gcggcgacgg caagtccggc aagatccaga ccgtgtgctt tgagggcgtg ctgaccatca
     2521 acgatgcccc agccctgatc gacctggtgc agcagggaat cggacctgct aagtcaatgg
     2581 gatgtgggct gctgtcactg gcacctctga agcggactgc tgatggcagt gaatttgagt
     2641 cctctagacc aaagaagaag agaaaggtgg gctccggcga gggcagggga agtcttctaa
     2701 catgcgggga cgtggaggaa aatcccggcc caatgccaaa gaagaagaga aaggtgggat
     2761 ccgaggagcc cttcaagtac atctgccact attggggcaa gagctccaag agcctgacaa
     2821 agggcaacga tatccacctg ctgatctacc actgcctgga cgtggcagca gtggcagatt
     2881 gttggtggga ccagtccgtg gtgctgcaga ataccttttg tcggaacgag atgctgtcta
     2941 agcagagagt gaaggcctgg ctgctgttct ttatcgccct gcacgatatc ggcaagttcg
     3001 acatccggtt tcagtataag tccgccgagt cttggctgaa gctgaatcca gccacaccct
     3061 ctctgaacgg ccctagcacc cagatgtgca gaaagttcaa tcacggcgcc gccggcctgt
     3121 actggtttaa ccaggactct ctgagcgagc agagcctggg cgatttcttt tccttctttg
     3181 acgcagcacc tcacccctac gagtcctggt tcccttgggt ggaggcagtg accggacacc
     3241 acggctttat cctgcattct caggaccagg ataagagccg gtgggagatg ccagccagcc
     3301 tggcctccta cgccgcacag gataagcagg ccagagagga gtggatcagc gtgctggagg
     3361 ccctgttcct gaccccagca ggactgtcca tcaatgatat cccacctgac tgctctagcc
     3421 tgctggccgg cttttgttcc ctggccgact ggctgggctc ttggaccaca accaatacat
     3481 tcctgtttaa cgaggatgca ccaagcgaca tcaacgccct gaggacctac ttccaggatc
     3541 ggcagcagga cgcctccaga gtgctggagc tgtccggcct ggtgtctaat aagaggtgct
     3601 acgagggagt gcacgccctg ctggataacg gctatcagcc aaggcagctg caggtgctgg
     3661 tggacgccct gcctgtggca ccaggactga cagtgatcga ggcacctacc ggcagcggca
     3721 agacagagac cgccctggcc tatgcctgga agctgatcga tcagcagatc gccgactccg
     3781 tgatcttcgc cctgccaaca caggccaccg ccaatgcaat gctgacccgg atggaggcct
     3841 ccgcctctca cctgttttcc tctcccaacc tgatcctggc ccacggcaac agcaggttca
     3901 atcacctgtt tcagtctatc aagagccgcg ccatcacaga gcagggacag gaggaggcct
     3961 gggtgcagtg ctgtcagtgg ctgagccagt ccaataagaa ggtgttcctg ggccagatcg
     4021 gcgtgtgcac catcgatcag gtgctgatct ctgtgctgcc tgtgaagcac aggtttatca
     4081 ggggactggg aatcggcaga agcgtgctga tcgtggatga ggtgcacgcc tacgacacat
     4141 atatgaacgg cctgctggag gccgtgctga aggcacaggc agacgtgggc ggctccgtga
     4201 tcctgctgtc tgccaccctg ccaatgaagc agaagcagaa gctgctggat acatacggcc
     4261 tgcacaccga ccccgtggag aacaattccg cctatcctct gatcaattgg aggggagtga
     4321 acggagcaca gcggttcgac ctgctggccc accctgagca gctgccacca aggttttcta
     4381 tccagcccga gcctatctgc ctggccgata tgctgccaga cctgaccatg ctggagagga
     4441 tgatcgctgc cgccaatgca ggagcacagg tgtgcctgat ctgtaacctg gtggatgtgg
     4501 cccaggtgtg ctaccagagg ctgaaggagc tgaacaatac acaggtggac atcgatctgt
     4561 tccacgcccg gtttaccctg aatgaccgga gagagaagga gaacagagtg atctctaact
     4621 tcggcaagaa tggcaagaga aacgtgggca gaatcctggt ggccacacag gtggtggagc
     4681 agagcctgga cgtggatttc gactggctga tcacccagca ctgccccgcc gatctgctgt
     4741 ttcagcggct gggcagactg cacaggcacc accgcaagta tcggcctgcc ggctttgaga
     4801 tcccagtggc cacaatcctg ctgcctgacg gagagggata cggcaggcac gagcacatct
     4861 atagcaatgt gcgcgtgatg tggcggaccc agcagcacat cgaggagctg aacggcgcct
     4921 ctctgttctt tccagatgcc taccgccagt ggctggacag catctatgac gatgccgaga
     4981 tggatgagcc cgagtgggtg ggcaatggca tggacaagtt cgagtccgcc gagtgtgaga
     5041 agcggttcaa ggccaggaag gtgctgcagt gggccgagga gtactccctg caggataacg
     5101 acgagacaat cctggccgtg accagagatg gcgagatgtc tctgcccctg ctgccttatg
     5161 tgcagacaag ctccggcaag cagctgctgg atggccaggt gtacgaggac ctgagccacg
     5221 agcagcagta tgaggccctg gccctgaaca gggtgaatgt gcccttcacc tggaagcgga
     5281 gcttcagcga agtggtggat gaggacggcc tgctgtggct ggagggcaag cagaatctgg
     5341 atggctgggt gtggcagggc aactccatcg tgatcacata caccggcgac gagggcatga
     5401 ccagagtgat cccagccaac cccaagaaga gaaccgccga cggcagcgag ttcgagtccg
     5461 ctagcccaaa gaagaagcgg aaggtctaaa agggttcgat ccctaccggt tagtaatgag
     5521 tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5581 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     5641 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5701 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5761 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgaa atcggatcca
     5821 gctgaatcta agtggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     5881 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     5941 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     6001 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     6061 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     6121 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     6181 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     6241 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     6301 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatctaag
     6361 ctgtgaccgg cgcctactct agagccgcca tgtctagact ggacaagagc aaagtcataa
     6421 actctgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     6481 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     6541 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccactcc tgccccctgg
     6601 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     6661 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     6721 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     6781 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     6841 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     6901 aacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     6961 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     7021 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     7081 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc ggggagggca
     7141 gaggaagtct actaacatgc ggtgacgtgg aggagaatcc cggccctatg gccaagcctt
     7201 tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca
     7261 tctctgagga ctacagcgtc gccagcgcag ctctctctag cgacggccgt atcttcactg
     7321 gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg
     7381 ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg
     7441 gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca
     7501 aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc
     7561 cctctggtta tgtgtgggag ggctgagtcg acaatcaacc tctggattac aaaatttgtg
     7621 ggttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     7681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     7741 atcttatcat gtctggaatt gactcaaatg atgtcaatta gtctatcaga agctatctgg
     7801 tctcccttcc gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact
     7861 gggttcttat atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa
     7921 cgaagtccct aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt
     7981 tttagggccg cggcccattg gtatggcttt ttccccgtat ccccccaggt gtctgcaggc
     8041 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     8101 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     8161 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     8221 ttgggggggg gctgtccctg atatctataa caagaaaata tatatataat aagttatcac
     8281 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     8341 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     8401 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     8461 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     8521 caattttacg cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg
     8581 gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga
     8641 agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg
     8701 ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc
     8761 catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg
     8821 gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac
     8881 cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg
     8941 atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc
     9001 ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     9061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     9121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     9181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     9361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     9421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     9481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     9541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     9601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     9661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     9721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     9781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     9841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     9901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     9961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    10021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    10081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    10141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    10201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    10261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    10381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    10441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    10501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    10561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    10621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    10681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    10741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    10801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    10861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    10921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    10981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    11041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    11101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    11161 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
    11221 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
    11281 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
    11341 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
    11401 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
    11461 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
    11521 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
    11581 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
    11641 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
    11701 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
    11761 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcctcgttca
    11821 ttcacgtttt tgaacccgtg gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg
    11881 tgatggagca gatgaagatg ctcgacacgc tgcagaacac gcagctaga
//
