LOCUS       sequence-308569-       10163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-Cutting and Integrating Payload CRISPR backbone with BFP 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10163)
  AUTHORS   Bloemberg D, Sosa-Miranda CD, Nguyen T, Weeratna RD, McComb S
  TITLE     Self-Cutting and Integrating CRISPR Plasmids Enable Targeted Genomic
            Integration of Genetic Payloads for Rapid Cell Engineering.
  JOURNAL   CRISPR J. 2021 Feb;4(1):104-119. doi: 10.1089/crispr.2020.0090.
  PUBMED    33616439
REFERENCE   2  (bases 1 to 10163)
  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"; doi: 
            "10.1089/crispr.2020"; journalName: "CRISPR J"; date: "2021-02"; 
            volume: "4"; issue: "1"; pages: "104-119"
COMMENT     Sequence Label: pSCIPpay-BFP reporter
FEATURES             Location/Qualifiers
     source          1..10163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             455..511
                     /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             512..1210
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     enhancer        1233..1518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1520..1797
                     /label=chicken beta-actin promoter
     intron          1798..2025
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2037..2046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2046..2111
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2118..2138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2163..3878
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     primer_bind     4360..4379
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6346..6393
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6409..6462
                     /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             join(6463..6465,6466..7176)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6463..6465,6466..7176)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6766..6785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7113..7134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7204..7221)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7210..7417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7426..7566
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7426..7555
                     /label=AAV2 ITR
     rep_origin      7641..8096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7728..7747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7938..7959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8113..8132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8232..8254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8292..8310)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8378..8482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8483..8551,8552..9343)
                     /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(8701..8720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9514..10102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10003..10022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctag agaccgtagg tctcaggcag cggcgccacc aacttcagcc tgctgaagca
      481 ggccggcgac gtggaggaga accccggccc cagcgagctg attaaggaga acatgcacat
      541 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
      601 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
      661 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
      721 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
      781 gagagtcacc acatacgagg acgggggcgt gctgaccgct acccaggaca ccagcctcca
      841 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
      901 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gaaacgctgt accccgctga
      961 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     1021 cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     1081 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg aaacctacgt
     1141 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     1201 gcttaattag taacagagac cgtaggtctc accgttacat aacttacggt aaatggcccg
     1261 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     1321 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1381 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1441 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1501 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     1561 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     1621 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     1681 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     1741 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     1801 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     1861 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     1921 ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta
     1981 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
     2041 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     2101 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     2161 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     2221 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     2281 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     2341 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2401 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2461 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2521 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2581 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2641 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2701 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2761 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     2821 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     2881 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     2941 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     3001 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     3061 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     3121 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     3181 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     3241 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3301 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3361 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3421 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3481 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3541 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3601 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3661 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3721 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3781 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     3841 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     3901 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     3961 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     4021 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     4081 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     4141 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     4201 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     4261 gcctgacctt taaagaggac atccagaaag cccaggtaag gggctcacag tagcaggctt
     4321 gaggtctgga catatatatg ggtgacaatg acatccactt tgcctttctc tccacaggtg
     4381 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     4441 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     4501 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     4561 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     4621 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     4681 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     4741 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     4801 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     4861 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     4921 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     4981 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     5041 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     5101 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     5161 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     5221 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     5281 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     5341 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     5401 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     5461 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     5521 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     5581 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     5641 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     5701 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     5761 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     5821 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     5881 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     5941 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     6001 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     6061 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     6121 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     6181 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     6241 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     6301 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
     6361 acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg gcagtggaga gggcagagga
     6421 agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag
     6481 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     6541 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     6601 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     6661 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     6721 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     6781 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     6841 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     6901 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     6961 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     7021 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     7081 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     7141 gccgggatca ctctcggcat ggacgagctg tacaaggaat tctaactaga gctcgctgat
     7201 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7261 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7321 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7381 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     7441 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     7501 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     7561 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7621 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     7681 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     7741 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     7801 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     7861 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7921 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     7981 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     8041 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     8101 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     8161 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     8221 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     8281 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     8341 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     8401 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     8461 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8521 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     8581 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     8641 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     8701 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     8761 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     8821 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8881 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8941 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     9001 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     9061 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     9121 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     9181 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     9241 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     9301 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     9361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     9421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     9481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
    10021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
    10081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
    10141 ttgctggcct tttgctcaca tgt
//
