LOCUS       sequence-307110-        9405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-Cutting and Integrating CRISPR backbone (px458 derivative with 
            Cas9-intron, sgRNA, and bait integration sites).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9405)
  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 9405)
  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: pQCi
FEATURES             Location/Qualifiers
     source          1..9405
                     /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"
     enhancer        475..760
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        762..1039
                     /label=chicken beta-actin promoter
     intron          1040..1267
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1279..1288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1288..1353
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1360..1380
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1405..3120
                     /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     3602..3621
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5588..5635
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5651..5704
                     /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(5705..5707,5708..6418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5705..5707,5708..6418)
                     /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(6008..6027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6355..6376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6446..6463)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6452..6659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6668..6808
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6668..6797
                     /label=AAV2 ITR
     rep_origin      6883..7338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6970..6989)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7180..7201
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7355..7374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7474..7496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7534..7552)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7620..7724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7725..7793,7794..8585)
                     /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(7943..7962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8756..9344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9245..9264
                     /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 aaatggctct agaacgcgta ggccttgaga cctagggtct ctgctagcgg tacccgttac
      481 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      541 aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      601 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      661 tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac
      721 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
      781 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
      841 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc crgggggggg
      901 gggggggggs ggggggsggg gsggggsgag gcggagaggt gcggcggcag ccaatcagag
      961 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     1021 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
     1081 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1141 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
     1201 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
     1261 ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat
     1321 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
     1381 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
     1441 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     1501 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     1561 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     1621 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     1681 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     1741 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     1801 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     1861 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1921 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1981 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     2041 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     2101 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     2161 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     2221 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     2281 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     2341 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     2401 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     2461 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     2521 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     2581 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     2641 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     2701 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     2761 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     2821 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     2881 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2941 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     3001 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     3061 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     3121 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     3181 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     3241 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     3301 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     3361 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     3421 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     3481 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggta
     3541 aggggctcac agtagcaggc ttgaggtctg gacatatata tgggtgacaa tgacatccac
     3601 tttgcctttc tctccacagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     3661 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     3721 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     3781 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     3841 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     3901 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     3961 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     4021 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     4081 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4141 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     4201 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     4261 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     4321 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     4381 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     4441 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     4501 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     4561 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     4621 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     4681 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     4741 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     4801 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     4861 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     4921 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     4981 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     5041 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     5101 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     5161 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     5221 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     5281 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     5341 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     5401 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     5461 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     5521 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     5581 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt
     5641 cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     5701 cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     5761 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     5821 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     5881 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     5941 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     6001 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     6061 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     6121 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     6181 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     6241 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     6301 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     6361 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagga
     6421 attctaacta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6481 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6541 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6601 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
     6661 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     6721 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     6781 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     6841 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     6901 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     6961 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7021 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7081 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7141 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7201 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     7261 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     7321 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7381 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7441 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7501 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7561 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     7621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag
     8461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8641 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8701 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8761 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8821 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8881 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8941 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9001 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9061 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9121 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9181 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9241 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9301 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9361 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
