LOCUS       sequence-307101-        9408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pQCi backbone with sgRNA targeting human TCR-alpha common chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9408)
  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 9408)
  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-hTRAC-sgRNA
FEATURES             Location/Qualifiers
     source          1..9408
                     /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"
     enhancer        478..763
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        765..1042
                     /label=chicken beta-actin promoter
     intron          1043..1270
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1282..1291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1291..1356
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1363..1383
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1408..3123
                     /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     3605..3624
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5591..5638
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5654..5707
                     /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(5708..5710,5711..6421)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5708..5710,5711..6421)
                     /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(6011..6030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6358..6379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6449..6466)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6455..6662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6671..6811
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6671..6800
                     /label=AAV2 ITR
     rep_origin      6886..7341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6973..6992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7183..7204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7358..7377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7477..7499
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7537..7555)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7623..7727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7728..7796,7797..8588)
                     /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(7946..7965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8759..9347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9248..9267
                     /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 tctctcagct ggtacacggc gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaacgc gtaggccttg agacctaggg tctctgctag cggtacccgt
      481 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      541 gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      601 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      661 caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc
      721 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
      781 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
      841 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccrggggg
      901 gggggggggg gggggggggs ggggsggggs grggcggaga ggtgcggcgg cagccaatca
      961 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     1021 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     1081 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     1141 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
     1201 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
     1261 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
     1321 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     1381 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     1441 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     1501 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     1561 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     1621 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     1681 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     1741 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     1801 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     1861 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     1921 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     1981 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     2041 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     2101 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     2161 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     2221 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     2281 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     2341 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     2401 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     2461 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     2521 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     2581 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     2641 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     2701 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     2761 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     2821 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     2881 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     2941 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     3001 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     3061 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     3121 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     3181 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     3241 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     3301 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     3361 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     3421 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     3481 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     3541 gtaaggggct cacagtagca ggcttgaggt ctggacatat atatgggtga caatgacatc
     3601 cactttgcct ttctctccac aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     3661 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     3721 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     3781 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     3841 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     3901 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     3961 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     4021 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     4081 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     4141 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     4201 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     4261 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     4321 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     4381 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     4441 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     4501 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     4561 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     4621 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     4681 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     4741 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     4801 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     4861 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     4921 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     4981 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     5041 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     5101 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     5161 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     5221 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     5281 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     5341 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     5401 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     5461 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     5521 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     5581 gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga
     5641 attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     5701 tggcccagtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     5761 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     5821 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     5881 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     5941 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     6001 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     6061 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     6121 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     6181 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     6241 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     6301 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     6361 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     6421 ggaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6481 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6541 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6601 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     6661 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     6721 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     6781 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     6841 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     6901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6961 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     7021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     7081 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     7141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     7201 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     7261 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     7321 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     7381 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     7441 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     7501 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     7561 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     7621 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7681 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7741 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7801 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7861 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7921 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7981 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     8041 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     8101 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     8161 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     8221 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     8281 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     8341 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     8401 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     8461 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     8521 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     8581 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     8641 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8701 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8761 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8821 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8881 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8941 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     9001 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     9061 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     9121 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     9181 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     9241 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     9301 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     9361 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
