LOCUS       sequence-231660-        9263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-sgRNAdest_CMV-intron_hProCas9Flavi_T2A Mcherry_AmpR_ColE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9263)
  AUTHORS   Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA, 
            Savage DF
  TITLE     Profiling of engineering hotspots identifies an allosteric 
            CRISPR-Cas9 switch.
  JOURNAL   Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528.
  PUBMED    27136077
REFERENCE   2  (bases 1 to 9263)
  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"; journalName: "Nat 
            Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528."
COMMENT     Sequence Label: pBLO43.3_human ProCas9Flavi
FEATURES             Location/Qualifiers
     source          1..9263
                     /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"
     enhancer        450..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        737..1014
                     /label=chicken beta-actin promoter
     intron          1015..1243
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1255..1264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1267..1287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2305..4803
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             5425..5445
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5446..5493
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             5515..5568
                     /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             5569..6279
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5714..5732)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6304..6321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6310..6517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6526..6666
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6526..6655
                     /label=AAV2 ITR
     rep_origin      6741..7196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6828..6847)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7038..7059
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7213..7232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7332..7354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7392..7410)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7478..7582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7583..7651,7652..8443)
                     /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(7801..7820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8614..9202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9103..9122
                     /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 taagagctat gctggaaaca gcatagcaag
      301 tttaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      361 ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa
      421 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
      481 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      781 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      841 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      901 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      961 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     1021 gctgcgacgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     1081 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     1141 ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat
     1201 gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc
     1261 atggccccaa agaagaagcg gaaggtcagc gctaacccca tcaacgccag cggcgtggac
     1321 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     1381 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     1441 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     1501 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     1561 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     1621 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     1681 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     1741 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     1801 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     1861 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     1921 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     1981 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     2041 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     2101 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     2161 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     2221 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     2281 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     2341 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     2401 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     2461 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     2521 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     2581 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     2641 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     2701 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     2761 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     2821 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     2881 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     2941 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     3001 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     3061 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     3121 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     3181 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     3241 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     3301 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     3361 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     3421 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     3481 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     3541 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     3601 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     3661 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     3721 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     3781 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     3841 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     3901 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     3961 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     4021 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     4081 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     4141 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     4201 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     4261 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     4321 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     4381 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     4441 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     4501 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     4561 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     4621 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     4681 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     4741 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     4801 gacaaacaaa agaaacgcgg aggcaaggac aagaagtaca gcatcggcct ggacatcggc
     4861 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     4921 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     4981 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     5041 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     5101 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     5161 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     5221 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     5281 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     5341 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     5401 aaccagctgt tcgaggaaac tagtccaaaa aagaagagaa aggtagatta caaagatgac
     5461 gatgacaaag actacaagga tgatgatgat aagggatccg aatccggcag tggagagggc
     5521 agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat ggtgagcaag
     5581 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     5641 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     5701 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     5761 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     5821 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     5881 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     5941 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaaa
     6001 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     6061 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     6121 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     6181 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     6241 cgccactcca ccggcggcat ggacgagctg tacaagtaag aattcctaga gctcgctgat
     6301 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6361 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6421 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6481 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     6541 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     6601 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     6661 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6721 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6781 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6841 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6901 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     6961 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7021 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     7081 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     7141 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     7201 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     7261 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7321 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7381 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7441 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7501 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7561 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7621 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7681 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7741 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7801 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7861 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7921 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7981 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8041 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     8101 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8161 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8221 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8281 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     8341 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8401 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8761 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9241 ttgctggcct tttgctcaca tgt
//
