LOCUS       sequence-220976-       13730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-sgRNA-EFS-arC9-T2A-mCherry-P2A-Hygro; arC9 without NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13730)
  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 13730)
  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: pBC2103-reci
FEATURES             Location/Qualifiers
     source          1..13730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1807..1924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1953..2201
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1953..1973
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2124..2143
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2376..2587
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2610..2633
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5010..7508
                     /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             7530..7583
                     /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             7584..8291
                     /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(7729..7747)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             8307..8363
                     /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"
     CDS             8364..9389
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    9425..10013
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9478..9498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9896..9907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10220..10453
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10525..10646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10562..10581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10616..10635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10686..10821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10868..10887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10869..10887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10897..10914)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10897..10913)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10906..10928)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11058..11513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11145..11164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11355..11376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11542..11646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11647..11715,11716..12507)
                     /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(11865..11884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12678..13266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13167..13186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13420..13437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13554..13575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13590..13595,13596..13613,13614..13620)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13628..13644
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     13633..13655
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13652..13668
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13652..13668
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13687..13707
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13689..13707
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacaataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     1921 ttttatcgat aacgagacta gcctcgatcg ccgagggcct atttcccatg attccttcat
     1981 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     2041 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     2101 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     2161 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggagacgga ggacgacgaa
     2221 cgtctctgtt taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt
     2281 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gaattcgcta gctaggtctt
     2341 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2401 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     2461 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2521 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2581 aacacaggac cggtagcgct cccgggatgg attacaaaga cgatgacgat aaggacaaga
     2641 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     2701 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     2761 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     2821 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     2881 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     2941 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     3001 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     3061 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     3121 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     3181 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     3241 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     3301 atctgatcgc ccagctgccc ggcgcatctc cagcaaacag tcttgccttg agtctcacag
     3361 cagaccagat ggtgtcagca ctgctcgatg cggagccacc tatactctac tctgaatacg
     3421 acccaaccag accgttcagc gaggcttcaa tgatgggact cctgactaat ctggcggaca
     3481 gggaacttgt gcacatgatc aattgggcca aacgggttcc cggttttgtt gaccttaccc
     3541 tgcatgatca ggttcatctg ttggaatgtg catggctcga aatacttatg attgggctcg
     3601 tatggaggtc catggagcac cctggcaagc tcctgtttgc tccaaatctc ttgttggatc
     3661 ggaatcaggg taaatgtgtc gaaggaatgg tagagatttt tgatatgctc ctggcaacct
     3721 ccagtcggtt ccgaatgatg aatctgcagg gggaggagtt cgtgtgtctg aaatctatta
     3781 tcctgcttaa ttccggcgtt tatacttttc tgtcatctac actgaagtcc ctggaggaga
     3841 aggaccacat tcatcgcgtg ttggacaaga taaccgacac actcatccat ctgatggcta
     3901 aggcagggct gaccctgcaa cagcaacacc agaggcttgc ccaactgttg ctcattctga
     3961 gccacatcag gcatatgtcc aataaaggaa tggaacacct gtacagcatg aagtgtaaga
     4021 acgtagtgcc actctatgat cttcttttgg agatgctgga cgctcacagg ctccacgcac
     4081 cacctgcatc tccaggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc
     4141 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     4201 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     4261 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     4321 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     4381 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     4441 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     4501 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     4561 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     4621 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     4681 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     4741 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     4801 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     4861 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     4921 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     4981 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     5041 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     5101 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     5161 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     5221 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     5281 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     5341 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     5401 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     5461 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     5521 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     5581 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     5641 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     5701 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     5761 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     5821 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     5881 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     5941 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     6001 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     6061 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     6121 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     6181 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     6241 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     6301 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     6361 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     6421 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     6481 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     6541 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     6601 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     6661 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     6721 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     6781 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     6841 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     6901 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     6961 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     7021 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     7081 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     7141 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     7201 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     7261 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     7321 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     7381 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     7441 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     7501 gaggcgacgg atccgaatcc ggcagtggag agggcagagg aagtctgcta acatgcggtg
     7561 acgtcgagga gaatcctggc ccaatggtga gcaagggcga ggaggataac atggccatca
     7621 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     7681 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     7741 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     7801 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     7861 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     7921 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     7981 acttcccctc cgacggcccc gtaatgcaga agaaaaccat gggctgggag gcctcctccg
     8041 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     8101 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     8161 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     8221 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     8281 agctgtacaa gcccggggga agcggagcta ctaacttcag cctgctgaag caggctggag
     8341 atgtggagga gaaccctgga cctatgaaaa agcctgaact caccgcgacg tctgtcgaga
     8401 agtttctgat cgaaaagttc gacagcgtat ccgacctgat gcagctctcg gagggcgaag
     8461 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     8521 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     8581 cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc
     8641 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     8701 agccggtcgc ggaggcaatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     8761 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     8821 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     8881 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     8941 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     9001 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     9061 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     9121 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     9181 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     9241 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     9301 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     9361 ccagcactcg tccgagggca aaggaatagg aattctaatg tcgacttcga aacgcgtctg
     9421 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     9481 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     9541 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     9601 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     9661 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     9721 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     9781 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     9841 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     9901 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     9961 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
    10021 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
    10081 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
    10141 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
    10201 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
    10261 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
    10321 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
    10381 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
    10441 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
    10501 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
    10561 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    10621 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
    10681 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
    10741 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
    10801 gaggaggctt ttttggaggc ctagggacgt acccaattcg cttttgcgtc gaaacgtacc
    10861 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
    10921 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
    10981 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
    11041 gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    11101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
    11161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
    11221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
    11281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    11341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    11401 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
    11461 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcccaggt
    11521 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
    11581 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
    11641 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
    11701 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
    11761 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
    11821 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
    11881 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
    11941 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
    12001 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
    12061 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
    12121 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
    12181 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
    12241 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
    12301 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
    12361 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
    12421 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
    12481 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    12541 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
    12601 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    12661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    12721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    12781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
    12841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    12901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    12961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    13021 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    13081 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    13141 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    13201 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    13261 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    13321 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
    13381 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
    13441 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
    13501 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
    13561 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
    13621 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
    13681 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
//
