LOCUS       sequence-256369-       11217 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Piggybac vector for constitutive expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11217)
  AUTHORS   Fischer AD, Veronese-Paniagua DA, Swaminathan S, Kashima H, Rubin 
            DC, Madison BB
  TITLE     The oncogenic function of PLAGL2 is mediated via ASCL2 and IGF2 and 
            a Wnt-independent mechanism in colorectal cancer.
  JOURNAL   Am J Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022.
  PUBMED    37310750
REFERENCE   2  (bases 1 to 11217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-ChPtW-eSpCas9
FEATURES             Location/Qualifiers
     source          1..11217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1875..1928
                     /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             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1974..1994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2019..4556,4557..4559,4560..5021,5022..5024,5025..5192,
                     5193..5195,5196..6119)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6120..6167
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    6194..6782
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6247..6267)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6665..6676)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6862..6996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6886..6905)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6940..6959
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7774..7836
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7966..7983)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7966..7982)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7975..7997)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8124..8579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8211..8230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8421..8442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8605..8709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8710..8778,8779..9570)
                     /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(8928..8947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9741..10329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10230..10249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10483..10500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10617..10638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10653..10658,10659..10676,10677..10683)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10691..10707
                     /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)."
     primer_bind     10696..10718
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10715..10731
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10715..10731
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      601 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      661 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
      721 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
      781 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
      841 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
      901 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
      961 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1021 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1081 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1141 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1201 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1261 ccatgactga atacaaacca actgttcgcc tggcaactcg tgatgatgtt ccacgtgcag
     1321 ttcgcaccct ggctgctgca tttgctgact accctgcaac ccgtcacact gtggacccag
     1381 accgccacat tgaacgtgtg actgaactgc aggagctgtt cctgacccgt gtgggcctgg
     1441 acattggcaa agtgtgggtg gcagatgatg gtgctgctgt ggcagtgtgg accacccctg
     1501 aatctgttga agctggtgca gtgtttgctg agattggccc acgcatggca gaactgtctg
     1561 gcagccgcct ggcagcacaa cagcagatgg aaggtctgct ggcaccacac cgcccaaaag
     1621 aacctgcttg gttcctggca actgtgggtg tgagccctga ccaccagggt aagggcctgg
     1681 gctctgcagt ggtgctgcct ggtgtggaag cagctgaacg tgcaggtgtg cctgctttcc
     1741 tggagacctc agctccacgc aacctgcctt tctatgaacg cctgggcttc actgtgactg
     1801 ctgatgtgga agtgccagaa ggcccacgca cttggtgcat gactcgcaaa ccaggtgcta
     1861 gccttggctc cggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     1921 ccggcccact cggctacccc tacgacgtgc ccgactacgc cagcggcatg gccccaaaga
     1981 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
     2041 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     2101 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     2161 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     2221 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     2281 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     2341 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     2401 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     2461 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     2521 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     2581 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     2641 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     2701 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     2761 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     2821 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     2881 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     2941 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     3001 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     3061 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     3121 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     3181 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     3241 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     3301 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     3361 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     3421 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     3481 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     3541 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     3601 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     3661 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     3721 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     3781 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     3841 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     3901 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     3961 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     4021 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     4081 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     4141 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     4201 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     4261 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     4321 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     4381 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     4441 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     4501 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctggccg
     4561 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     4621 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     4681 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     4741 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     4801 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     4861 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     4921 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     4981 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc tgcgctggaa agcgagttcg
     5041 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     5101 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     5161 agattaccct ggccaacggc gagatccgga aggcgcctct gatcgagaca aacggcgaaa
     5221 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     5281 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     5341 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     5401 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     5461 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     5521 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     5581 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     5641 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     5701 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     5761 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     5821 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     5881 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     5941 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     6001 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     6061 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     6121 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gagctagagt
     6181 tcgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6241 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     6301 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6361 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6421 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6481 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6541 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     6601 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6661 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6721 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6781 gcctggaatt aattctggct aagcacttcg tggccgtcga tcgtttaaag ggaggtagtg
     6841 agtcgagata tcgatactag tgatccagac atgataagat acattgatga gtttggacaa
     6901 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     6961 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     7021 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     7081 tgtggtatgg ctgattatga tcctctagag ggacagcccc cccccaaagc ccccagggat
     7141 gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct ccgctccggt
     7201 ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg gcacggggaa
     7261 ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc
     7321 tggggggata cggggaaaag ccataccaat ggagctcacg gggacagccc ccccccaaag
     7381 cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc
     7441 tccgctccgg tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg
     7501 ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc
     7561 ctgcagacac ctggggggat acggggaaaa agaattcgat aaaagttttg ttactttata
     7621 gaagaaattt tgagtttttg ttttttttaa taaataaata aacataaata aattgtttgt
     7681 tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa
     7741 taaataaacc tcgatataca gaccgataaa acacatgcgt caattttaca catgattatc
     7801 tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagctgcg tgttctgcag
     7861 cgtgtcgagc atcttcatct gctccatcac gctgtaaaac acatttgcac cgcgagtctg
     7921 cccgtcctcc acgggttcaa aaacgtgaat gaacgaggcg cgctcactgg ccgtcgtttt
     7981 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     8041 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     8101 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     8161 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     8221 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     8281 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     8341 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     8401 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     8461 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     8521 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     8581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     8641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     8701 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     8761 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     8821 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     8881 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     8941 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     9001 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     9061 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     9121 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     9181 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     9241 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     9301 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     9361 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     9421 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     9481 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     9541 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     9601 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     9661 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     9721 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     9781 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     9841 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     9901 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     9961 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
    10021 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
    10081 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
    10141 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
    10201 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
    10261 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
    10321 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
    10381 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
    10441 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
    10501 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
    10561 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
    10621 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
    10681 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
    10741 gccaagcgcg cccgccgggt aactcacggg gtatccatgt ccatttctgc ggcatccagc
    10801 caggataccc gtcctcgctg acgtaatatc ccagcgccgc accgctgtca ttaatctgca
    10861 caccggcacg gcagttccgg ctgtcgccgg tattgttcgg gttgctgatg cgcttcgggc
    10921 tgaccatccg gaactgtgtc cggaaaagcc gcgacgaact ggtatcccag gtggcctgaa
    10981 cgaacagttc accgttaaag gcgtgcatgg ccacaccttc ccgaatcatc atggtaaacg
    11041 tgcgttttcg ctcaacgtca atgcagcagc agtcatcctc ggcaaactct ttccatgccg
    11101 cttcaacctc gcgggaaaag gcacgggctt cttcctcccc gatgcccaga tagcgccagc
    11161 ttgggcgatg actgagccgg aaaaaagacc cgacgatatg atcctgatgc agctaga
//
