LOCUS       sequence-373707-       11883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged SaCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11883)
  AUTHORS   Escobar M, Li J, Patel A, Liu S, Xu Q, Hilton IB
  TITLE     Quantification of Genome Editing and Transcriptional Control 
            Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in
            Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3239-3250. doi: 
            10.1021/acssynbio.2c00156. Epub 2022 Sep 26.
  PUBMED    36162812
REFERENCE   2  (bases 1 to 11883)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.2c00156"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3239-3250"
COMMENT     Sequence Label: SaCas9
FEATURES             Location/Qualifiers
     source          1..11883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        2071..2146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2208..2419
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2433..5591
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             5601..5648
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5655..5678
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5688..5744
                     /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             5745..6341
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6238..6258
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6357..6945
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6410..6430)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6828..6839)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7017..7250
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7276..7293)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7282..7506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7552..7980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7639..7658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7849..7870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7989..8009)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7994..8323
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8174..8309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8236..8255
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8371..8418
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8437..8811
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8941..9062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8978..8997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9032..9051
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9111..9127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9111..9127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9124..9146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9135..9151
                     /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)."
     promoter        complement(join(9159..9165,9166..9183,9184..9189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9204..9225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9342..9359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9513..10101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9593..9612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10272..11063,11064..11132))
                     /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     10895..10914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11133..11237)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11312..11331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11503..11882
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg atatcactag tgttccatta actgcgctaa aaataatttt taaatctttt
     2041 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2101 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     2161 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2221 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2281 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2341 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2401 gtttgccgcc agaacacagg accggtgcca ccatgaaaag gaactacatt ctggggctgg
     2461 acatcgggat tacaagcgtg gggtatggga ttattgacta tgaaacaagg gacgtgatcg
     2521 acgcaggcgt cagactgttc aaggaggcca acgtggaaaa caatgaggga cggagaagca
     2581 agaggggagc caggcgcctg aaacgacgga gaaggcacag aatccagagg gtgaagaaac
     2641 tgctgttcga ttacaacctg ctgaccgacc attctgagct gagtggaatt aatccttatg
     2701 aagccagggt gaaaggcctg agtcagaagc tgtcagagga agagttttcc gcagctctgc
     2761 tgcacctggc taagcgccga ggagtgcata acgtcaatga ggtggaagag gacaccggca
     2821 acgagctgtc tacaaaggaa cagatctcac gcaatagcaa agctctggaa gagaagtatg
     2881 tcgcagagct gcagctggaa cggctgaaga aagatggcga ggtgagaggg tcaattaata
     2941 ggttcaagac aagcgactac gtcaaagaag ccaagcagct gctgaaagtg cagaaggctt
     3001 accaccagct ggatcagagc ttcatcgata cttatatcga cctgctggag actcggagaa
     3061 cctactatga gggaccagga gaagggagcc ccttcggatg gaaagacatc aaggaatggt
     3121 acgagatgct gatgggacat tgcacctatt ttccagaaga gctgagaagc gtcaagtacg
     3181 cttataacgc agatctgtac aacgccctga atgacctgaa caacctggtc atcaccaggg
     3241 atgaaaacga gaaactggaa tactatgaga agttccagat catcgaaaac gtgtttaagc
     3301 agaagaaaaa gcctacactg aaacagattg ctaaggagat cctggtcaac gaagaggaca
     3361 tcaagggcta ccgggtgaca agcactggaa aaccagagtt caccaatctg aaagtgtatc
     3421 acgatattaa ggacatcaca gcacggaaag aaatcattga gaacgccgaa ctgctggatc
     3481 agattgctaa gatcctgact atctaccaga gctccgagga catccaggaa gagctgacta
     3541 acctgaacag cgagctgacc caggaagaga tcgaacagat tagtaatctg aaggggtaca
     3601 ccggaacaca caacctgtcc ctgaaagcta tcaatctgat tctggatgag ctgtggcata
     3661 caaacgacaa tcagattgca atctttaacc ggctgaagct ggtcccaaaa aaggtggacc
     3721 tgagtcagca gaaagagatc ccaaccacac tggtggacga tttcattctg tcacccgtgg
     3781 tcaagcggag cttcatccag agcatcaaag tgatcaacgc catcatcaag aagtacggcc
     3841 tgcccaatga tatcattatc gagctggcta gggagaagaa cagcaaggac gcacagaaga
     3901 tgatcaatga gatgcagaaa cgaaaccggc agaccaatga acgcattgaa gagattatcc
     3961 gaactaccgg gaaagagaac gcaaagtacc tgattgaaaa aatcaagctg cacgatatgc
     4021 aggagggaaa gtgtctgtat tctctggagg ccatccccct ggaggacctg ctgaacaatc
     4081 cattcaacta cgaggtcgat catattatcc ccagaagcgt gtccttcgac aattccttta
     4141 acaacaaggt gctggtcaag caggaagaga actctaaaaa gggcaatagg actcctttcc
     4201 agtacctgtc tagttcagat tccaagatct cttacgaaac ctttaaaaag cacattctga
     4261 atctggccaa aggaaagggc cgcatcagca agaccaaaaa ggagtacctg ctggaagagc
     4321 gggacatcaa cagattctcc gtccagaagg attttattaa ccggaatctg gtggacacaa
     4381 gatacgctac tcgcggcctg atgaatctgc tgcgatccta tttccgggtg aacaatctgg
     4441 atgtgaaagt caagtccatc aacggcgggt tcacatcttt tctgaggcgc aaatggaagt
     4501 ttaaaaagga gcgcaacaaa gggtacaagc accatgccga agatgctctg attatcgcaa
     4561 atgccgactt catctttaag gagtggaaaa agctggacaa agccaagaaa gtgatggaga
     4621 accagatgtt cgaagagaag caggccgaat ctatgcccga aatcgagaca gaacaggagt
     4681 acaaggagat tttcatcact cctcaccaga tcaagcatat caaggatttc aaggactaca
     4741 agtactctca ccgggtggat aaaaagccca acagagagct gatcaatgac accctgtata
     4801 gtacaagaaa agacgataag gggaataccc tgattgtgaa caatctgaac ggactgtacg
     4861 acaaagataa tgacaagctg aaaaagctga tcaacaaaag tcccgagaag ctgctgatgt
     4921 accaccatga tcctcagaca tatcagaaac tgaagctgat tatggagcag tacggcgacg
     4981 agaagaaccc actgtataag tactatgaag agactgggaa ctacctgacc aagtatagca
     5041 aaaaggataa tggccccgtg atcaagaaga tcaagtacta tgggaacaag ctgaatgccc
     5101 atctggacat cacagacgat taccctaaca gtcgcaacaa ggtggtcaag ctgtcactga
     5161 agccatacag attcgatgtc tatctggaca acggcgtgta taaatttgtg actgtcaaga
     5221 atctggatgt catcaaaaag gagaactact atgaagtgaa tagcaagtgc tacgaagagg
     5281 ctaaaaagct gaaaaagatt agcaaccagg cagagttcat cgcctccttt tacaacaacg
     5341 acctgattaa gatcaatggc gaactgtata gggtcatcgg ggtgaacaat gatctgctga
     5401 accgcattga agtgaatatg attgacatca cttaccgaga gtatctggaa aacatgaatg
     5461 ataagcgccc ccctcgaatt atcaaaacaa ttgcctctaa gactcagagt atcaaaaagt
     5521 actcaaccga cattctggga aacctgtatg aggtgaagag caaaaagcac cctcagatta
     5581 tcaaaaaggg cagcggaggc aagcgtcctg ctgctactaa gaaagctggt caagctaaga
     5641 aaaagaaagg atcagattac aaagacgatg acgataaggg atccggcgca acaaacttct
     5701 ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgaccgag tacaagccca
     5761 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     5821 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     5881 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5941 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     6001 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     6061 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     6121 ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     6181 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     6241 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     6301 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc
     6361 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6421 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6481 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     6541 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     6601 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     6661 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     6721 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6781 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6841 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6901 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc
     6961 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     7021 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     7081 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7141 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7201 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
     7261 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     7321 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     7381 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     7441 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     7501 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     7561 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     7621 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7681 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     7741 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     7801 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7861 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     7921 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7981 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     8041 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     8101 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     8161 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     8221 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     8281 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     8341 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     8401 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     8461 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     8521 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     8581 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     8641 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     8701 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     8761 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     8821 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     8881 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     8941 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     9001 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     9061 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     9121 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     9181 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     9241 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     9301 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     9361 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     9421 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     9481 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     9541 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     9601 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     9661 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     9721 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     9781 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     9841 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     9901 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     9961 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    10021 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    10081 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    10141 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    10201 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    10261 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    10321 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    10381 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    10441 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    10501 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    10561 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    10621 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    10681 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    10741 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    10801 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    10861 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    10921 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    10981 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    11041 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    11101 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    11161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    11221 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
    11281 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
    11341 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
    11401 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
    11461 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
    11521 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
    11581 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
    11641 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
    11701 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
    11761 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
    11821 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
    11881 tgg
//
