LOCUS       sequence-307109-       12318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-Cutting and Integrating CRISPR backbone targeting human CD69 
            promoter with TdTomato reporter transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12318)
  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 12318)
  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: pSCIP-hCD69-TdTomato
FEATURES             Location/Qualifiers
     source          1..12318
                     /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        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1033..1089
                     /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"
     primer_bind     complement(1946..1964)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2408..2431
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2560..2767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2786..2802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3388..3673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3675..3952
                     /label=chicken beta-actin promoter
     intron          3953..4180
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4192..4201
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4201..4266
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4273..4293
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4318..6033
                     /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     6515..6534
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             8501..8548
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8564..8617
                     /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(8618..8620,8621..9331)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8618..8620,8621..9331)
                     /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(8921..8940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9268..9289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9365..9572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   9581..9721
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   9581..9710
                     /label=AAV2 ITR
     rep_origin      9796..10251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9883..9902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10093..10114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10268..10287)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10387..10409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10447..10465)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10533..10637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10638..10706,10707..11498)
                     /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(10856..10875)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11669..12257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12158..12177
                     /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 agctctttgc atccggagag gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaacgc gtaggcctag agataccaga gagtaaatag gttactttct
      481 tgtccactct ccggagctct ttgcatccgg agagtggtcc tggaaaatac tacactctcc
      541 ttctccaaat tatcttcatc tcaaagacag gaacctctaa cttttaattc tttacttaga
      601 ttatgctgtc tcctaaactg tttatgtttt ctagaaattt aaggcaggat gtctcagagt
      661 ctgggaaaat cccactttcc tcctgctaca ccttacagtt gtgagaaagc acatttcaga
      721 caacagggaa aacccatact tcaccacaac aacacactat acattgtctg gtccactgga
      781 gcataaatta aagagaaaca atgtagtcaa gcaagtaggc ggcaagagga agggggcgga
      841 gacatcatca gggagtataa actctgagat gcctcagagc ctcacagact caacaagagc
      901 tccagcaaag actttcactg tagcttgact tgacctgaga ttaactaggg aatcttgaga
      961 ataaagatga gctctgaaaa ttgtttcgta gcagagaaca gctctttgca tccggagctc
     1021 gagggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt ggaggagaac
     1081 cccggccccg ttagcaaagg tgaagaagta attaaggaat ttatgcgttt taaagttcgt
     1141 atggaaggat ctatgaatgg gcatgaattt gaaattgaag gagaaggtga aggacggcca
     1201 tatgaaggaa cacaaacagc taaactcaaa gtcactaaag gtggacctct cccttttgcg
     1261 tgggatattc tcagcccaca atttatgtat gggagcaaag cctatgtaaa acatccagct
     1321 gatattcctg actataagaa gctcagcttt cctgaaggat ttaaatggga acgtgtgatg
     1381 aattttgaag atggtggcct cgtcactgta acacaagatt catctttgca agatggtact
     1441 cttatttata aagtcaaaat gagagggact aactttccac cagatggacc agtcatgcaa
     1501 aagaaaacta tggggtggga agcttcaaca gaaagattgt atccacggga cggtgtattg
     1561 aaaggagaaa ttcatcaagc actcaaattg aaagacggtg gacattattt ggtcgaattt
     1621 aaaacaattt atatggctaa gaaaccagta cagctccctg gctattatta tgtcgatact
     1681 aaattggata ttacatcaca taacgaagat tatactatcg tggagcaata tgaacggagc
     1741 gaaggtagac atcatctgtt tctcggtcac ggtactggat ctacagggtc cggttctagc
     1801 ggaactgcta gctctgaaga taataatatg gcggtgatta aggaatttat gagatttaaa
     1861 gttcgaatgg aaggatctat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1921 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     1981 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     2041 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     2101 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     2161 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     2221 atgcagaaga aaaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     2281 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     2341 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     2401 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2461 cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaatag
     2521 taagtcgaca cgcgtctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     2581 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2641 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2701 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     2761 ctggggaaag cttggcgtaa tcatggtcat agctgtttcc tgctatagtg gacaagaaag
     2821 taagtcagat ctgattgctt atttactctc tggtatctct atatatgaca ttaatagtag
     2881 ttatacaaga tatactggta atctctctat atatgatata taatcttaga gaaatatact
     2941 gctaatctct atgtatgtga tatctaatgg ttgttaggga aggaagtgag gctttgtctt
     3001 ttagagagga caggttcttc ttcagggaag aggtaaaccc ctttttatct ataagaagat
     3061 agaatacctg agatgagaga aagccgcaaa aatgaatttc agaagatctt taaagatctt
     3121 tctctttgct ttagtttata tcatctcagt gtttagattt aattagaaaa ctttctgctc
     3181 tagcccagtt attctccatc atttttttca tatccatctg tacattctcc aggaagtgtt
     3241 gtctctcttt atgaaaaatt ctgttttcca cataagcaat tacattgttt gccttccttt
     3301 gcattcccac tctccggatg caaagagctc cggagagtgg acaagaaagt aacctattta
     3361 ctctctggta tctctgctag cggtacccgt tacataactt acggtaaatg gcccgcctgg
     3421 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     3481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3541 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     3601 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3661 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     3721 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     3781 gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsggggggs ggggsggggc
     3841 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     3901 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     3961 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     4021 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     4081 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
     4141 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
     4201 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
     4261 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
     4321 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
     4381 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     4441 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     4501 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     4561 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     4621 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     4681 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     4741 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     4801 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     4861 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     4921 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     4981 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     5041 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     5101 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     5161 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     5221 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     5281 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     5341 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     5401 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     5461 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     5521 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     5581 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     5641 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     5701 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     5761 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     5821 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     5881 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     5941 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     6001 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     6061 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     6121 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     6181 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     6241 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     6301 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     6361 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     6421 acctttaaag aggacatcca gaaagcccag gtaaggggct cacagtagca ggcttgaggt
     6481 ctggacatat atatgggtga caatgacatc cactttgcct ttctctccac aggtgtccgg
     6541 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     6601 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     6661 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     6721 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     6781 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     6841 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     6901 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     6961 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     7021 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     7081 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     7141 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     7201 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     7261 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     7321 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     7381 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     7441 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     7501 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     7561 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     7621 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     7681 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     7741 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     7801 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     7861 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     7921 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     7981 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     8041 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     8101 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     8161 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     8221 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     8281 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     8341 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     8401 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     8461 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa
     8521 aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca gaggaagtct
     8581 gctaacatgc ggtgacgtcg aggagaatcc tggcccagtg agcaagggcg aggagctgtt
     8641 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     8701 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     8761 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     8821 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     8881 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     8941 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     9001 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     9061 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     9121 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     9181 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     9241 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     9301 gatcactctc ggcatggacg agctgtacaa ggaattctaa ctagagctcg ctgatcagcc
     9361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     9421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     9481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     9541 gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag
     9601 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     9661 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     9721 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     9781 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     9841 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     9901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     9961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
    10021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
    10081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
    10141 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
    10201 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
    10261 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
    10321 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
    10381 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
    10441 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
    10501 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
    10561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
    10621 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
    10681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
    10741 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
    10801 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
    10861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
    10921 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
    10981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
    11041 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
    11101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
    11161 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
    11221 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
    11281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
    11341 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
    11401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
    11461 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
    11521 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
    11581 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
    11641 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    11701 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    11761 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    11821 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    11881 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    11941 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    12001 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    12061 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    12121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    12181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    12241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    12301 ggccttttgc tcacatgt
//
