LOCUS       sequence-224902-       13714 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 13714)
  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 13714)
  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..13714
                     /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"
     promoter        424..635
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             658..681
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3058..5556
                     /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             5578..5631
                     /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             5632..6339
                     /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(5777..5795)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             6355..6411
                     /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             6412..7437
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    7473..8061
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7526..7546)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7944..7955)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8268..8501
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8573..8694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8610..8629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8664..8683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8734..8869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8907..8926)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8908..8926)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8936..8953)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8936..8952)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8945..8967)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9094..9549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9181..9200)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9391..9412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9575..9679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9680..9748,9749..10540)
                     /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(9898..9917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10711..11299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11200..11219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11453..11470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11587..11608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11623..11628,11629..11646,11647..11653)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11661..11677
                     /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     11666..11688
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11685..11701
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11685..11701
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11720..11740
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11722..11740
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11768..11994
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11995..12175
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12222..12347
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12840..13073
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13258..13302
                     /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    13569..13686
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gagacggagg acgacgaacg tctctgttta agagctatgc tggaaacagc
      301 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      361 ctttttttga attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      421 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
      481 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
      541 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
      601 ttctttttcg caacgggttt gccgccagaa cacaggaccg gtagcgctcc cgggatggat
      661 tacaaagacg atgacgataa ggacaagaag tacagcatcg gcctggacat cggcaccaac
      721 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
      781 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
      841 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
      901 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
      961 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     1021 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     1081 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1141 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     1201 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     1261 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     1321 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgcatctcca
     1381 gcaaacagtc ttgccttgag tctcacagca gaccagatgg tgtcagcact gctcgatgcg
     1441 gagccaccta tactctactc tgaatacgac ccaaccagac cgttcagcga ggcttcaatg
     1501 atgggactcc tgactaatct ggcggacagg gaacttgtgc acatgatcaa ttgggccaaa
     1561 cgggttcccg gttttgttga ccttaccctg catgatcagg ttcatctgtt ggaatgtgca
     1621 tggctcgaaa tacttatgat tgggctcgta tggaggtcca tggagcaccc tggcaagctc
     1681 ctgtttgctc caaatctctt gttggatcgg aatcagggta aatgtgtcga aggaatggta
     1741 gagatttttg atatgctcct ggcaacctcc agtcggttcc gaatgatgaa tctgcagggg
     1801 gaggagttcg tgtgtctgaa atctattatc ctgcttaatt ccggcgttta tacttttctg
     1861 tcatctacac tgaagtccct ggaggagaag gaccacattc atcgcgtgtt ggacaagata
     1921 accgacacac tcatccatct gatggctaag gcagggctga ccctgcaaca gcaacaccag
     1981 aggcttgccc aactgttgct cattctgagc cacatcaggc atatgtccaa taaaggaatg
     2041 gaacacctgt acagcatgaa gtgtaagaac gtagtgccac tctatgatct tcttttggag
     2101 atgctggacg ctcacaggct ccacgcacca cctgcatctc caggcgagaa gaagaatggc
     2161 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     2221 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     2281 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     2341 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     2401 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     2461 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     2521 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     2581 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     2641 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     2701 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     2761 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     2821 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     2881 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     2941 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     3001 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     3061 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     3121 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     3181 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     3241 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     3301 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     3361 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     3421 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     3481 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     3541 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     3601 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     3661 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     3721 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     3781 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     3841 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     3901 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     3961 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     4021 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     4081 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     4141 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     4201 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     4261 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     4321 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     4381 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     4441 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     4501 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     4561 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     4621 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     4681 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     4741 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     4801 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     4861 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     4921 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     4981 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     5041 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     5101 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     5161 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     5221 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     5281 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     5341 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     5401 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     5461 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     5521 gagacacgga tcgacctgtc tcagctggga ggcgacggat ccgaatccgg cagtggagag
     5581 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatggtgagc
     5641 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     5701 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     5761 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     5821 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     5881 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     5941 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     6001 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     6061 aaaaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     6121 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     6181 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     6241 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     6301 ggccgccact ccaccggcgg catggacgag ctgtacaagc ccgggggaag cggagctact
     6361 aacttcagcc tgctgaagca ggctggagat gtggaggaga accctggacc tatgaaaaag
     6421 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtatcc
     6481 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     6541 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     6601 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggagttc
     6661 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     6721 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga tgcgatcgct
     6781 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     6841 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     6901 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     6961 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     7021 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     7081 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     7141 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     7201 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     7261 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     7321 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     7381 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataggaa
     7441 ttctaatgtc gacttcgaaa cgcgtctgga acaatcaacc tctggattac aaaatttgtg
     7501 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     7561 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     7621 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     7681 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     7741 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7801 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7861 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7921 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7981 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     8041 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg
     8101 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     8161 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     8221 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     8281 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     8341 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     8401 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     8461 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     8521 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     8581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     8701 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     8761 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     8821 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     8881 ccaattcgcg aaacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     8941 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     9001 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     9061 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     9121 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     9181 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     9241 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     9301 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     9361 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     9421 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     9481 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     9541 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     9601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     9661 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     9721 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     9781 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     9841 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     9901 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     9961 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
    10021 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
    10081 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
    10141 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
    10201 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
    10261 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
    10321 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
    10381 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
    10441 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
    10501 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
    10561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
    10621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    10681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    10741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    10801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    10861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    10921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    10981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    11041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    11101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    11161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    11221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    11281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    11341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    11401 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    11461 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    11521 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
    11581 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
    11641 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    11701 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
    11761 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
    11821 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
    11881 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
    11941 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
    12001 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
    12061 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
    12121 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
    12181 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
    12241 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
    12301 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
    12361 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
    12421 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
    12481 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
    12541 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
    12601 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
    12661 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
    12721 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
    12781 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
    12841 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
    12901 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
    12961 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
    13021 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
    13081 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
    13141 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
    13201 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
    13261 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
    13321 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
    13381 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
    13441 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
    13501 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
    13561 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
    13621 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
    13681 aattttatcg atcacgagac tagcctcgat cgcc
//
