LOCUS       sequence-307107-        9414 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Self-Cutting and Integrating CRISPR backbone targeting human 
            TCR-alpha common chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9414)
  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 9414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1089/crispr.2020"; journalName: "CRISPR J"; date: "2021-02"; 
            volume: "4"; issue: "1"; pages: "104-119"
COMMENT     Sequence Label: pSCIP-hTRAC
FEATURES             Location/Qualifiers
     source          1..9414
                     /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        484..769
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        771..1048
                     /label=chicken beta-actin promoter
     intron          1049..1276
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1288..1297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1297..1362
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1369..1389
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1414..3129
                     /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     3611..3630
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5597..5644
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5660..5713
                     /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(5714..5716,5717..6427)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5714..5716,5717..6427)
                     /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(6017..6036)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6364..6385
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6455..6472)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6461..6668
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6677..6817
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6677..6806
                     /label=AAV2 ITR
     rep_origin      6892..7347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6979..6998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7189..7210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7364..7383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7483..7505
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7543..7561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7629..7733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7734..7802,7803..8594)
                     /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(7952..7971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8765..9353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9254..9273
                     /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 gtaggccttc tctcagctgg tacacggcag ggctagcggt
      481 acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      541 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      601 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      661 tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga
      721 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      781 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      841 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      901 gggggggggg ggggggggsg gggggsgggg sggggsgagg cggagaggtg cggcggcagc
      961 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     1021 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     1081 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     1141 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg
     1201 gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga
     1261 aatcactttt tttcaggttg gaccggtgcc accatggact ataaggacca cgacggagac
     1321 tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag
     1381 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
     1441 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     1501 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     1561 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     1621 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     1681 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     1741 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     1801 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     1861 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     1921 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     1981 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2041 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2101 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     2161 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2221 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2281 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     2341 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     2401 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     2461 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     2521 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     2581 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     2641 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     2701 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     2761 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     2821 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     2881 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     2941 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3001 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3061 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3121 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3181 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3241 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3301 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     3361 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     3421 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     3481 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     3541 gcccaggtaa ggggctcaca gtagcaggct tgaggtctgg acatatatat gggtgacaat
     3601 gacatccact ttgcctttct ctccacaggt gtccggccag ggcgatagcc tgcacgagca
     3661 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     3721 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     3781 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     3841 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     3901 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     3961 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     4021 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     4081 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     4141 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     4201 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     4261 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     4321 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     4381 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     4441 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     4501 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     4561 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     4621 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     4681 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     4741 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     4801 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     4861 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     4921 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     4981 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     5041 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     5101 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     5161 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     5221 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     5281 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     5341 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     5401 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     5461 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     5521 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     5581 tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa
     5641 aaaggaattc ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga
     5701 gaatcctggc ccagtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     5761 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     5821 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     5881 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     5941 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     6001 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     6061 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     6121 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     6181 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     6241 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     6301 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     6361 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     6421 gtacaaggaa ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     6481 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6541 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     6601 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat
     6661 gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     6721 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     6781 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     6841 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     6901 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6961 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7021 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     7081 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     7141 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7201 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     7261 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7321 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     7381 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7441 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7501 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     7561 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     7621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7741 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7801 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7861 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7921 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7981 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     8041 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     8101 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     8161 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     8221 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     8281 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     8341 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     8401 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     8461 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8521 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8581 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8641 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8701 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8761 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8821 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8881 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     8941 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     9001 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     9061 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     9121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     9181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     9241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     9301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     9361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
