LOCUS       sequence-179756-        8859 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pAC154-dual-dCas9VP160-sgExpression with MYRF sg4 cloned into BbsI 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  AUTHORS   Pouladi M
  TITLE     Pouladi lab MYRF plasmids 2017
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8859)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: pYTP MYRF sg4
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..42
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             46..66
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(76..99,100..102,103..2658,2659..2661,2662..4176)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(76..99,100..102,103..2589,2590..2592,2593..4176)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4180..4200
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4234..4617
                     /codon_start=1
                     /product="10 tandem repeats of the minimal activation 
                     domain of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP160
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS
                     DALDDFDLDMLGSDALDDFDLDML"
     primer_bind     complement(4662..4679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4668..4875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4884..5024
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4884..5013
                     /label=AAV2 ITR
     rep_origin      5099..5554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5186..5205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5396..5417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5571..5590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5690..5712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5750..5768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5836..5940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5941..6009,6010..6801)
                     /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(6159..6178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6972..7560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7461..7480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7622..7870
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7622..7642
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7793..7812
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7891..7966
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8063..8348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        8350..8626
                     /label=chicken beta-actin promoter
     intron          8627..8854
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 accggtgcca ccatgtaccc atacgatgtt ccagattacg cttcgccgaa gaaaaagcgc
       61 aaggtcgaag cgtccgacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
      121 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
      181 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
      241 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
      301 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
      361 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
      421 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
      481 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
      541 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
      601 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
      661 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
      721 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
      781 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
      841 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
      901 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
      961 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     1021 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     1081 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     1141 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     1201 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     1261 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     1321 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     1381 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     1441 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     1501 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     1561 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     1621 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     1681 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     1741 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     1801 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     1861 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     1921 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     1981 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     2041 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     2101 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     2161 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     2221 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     2281 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     2341 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     2401 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     2461 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     2521 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     2581 gatgtggacg ccatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     2641 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     2701 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     2761 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     2821 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     2881 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     2941 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     3001 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     3061 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     3121 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     3181 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     3241 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     3301 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     3361 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     3421 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     3481 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     3541 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     3601 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     3661 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     3721 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     3781 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     3841 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     3901 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     3961 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     4021 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     4081 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     4141 gagacacgga tcgacctgtc tcagctggga ggcgacagcc ccaagaagaa gagaaaggtg
     4201 gaggccagcg ggccggccgg atccgggcgc gccgacgcgc tggacgattt cgatctcgac
     4261 atgctgggtt ctgatgccct cgatgacttt gacctggata tgttgggaag cgacgcattg
     4321 gatgactttg atctggacat gctcggctcc gatgctctgg acgatttcga tctcgatatg
     4381 ttagggtcag acgcactgga tgatttcgac cttgatatgt tgggaagcga tgcccttgat
     4441 gatttcgacc tggacatgct cggcagcgac gccctggacg atttcgatct ggacatgctg
     4501 gggtccgatg ccttggatga ttttgacttg gatatgctgg ggagtgatgc cctggacgac
     4561 tttgacctgg acatgctggg ctccgatgcg ctcgatgact tcgatttgga tatgttgtat
     4621 atcgattgat taattaagaa ttcctagagc tcgctgatca gcctcgactg tgccttctag
     4681 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4741 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4801 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     4861 caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4921 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4981 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     5041 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     5101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5161 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5221 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5281 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     5341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5401 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     5461 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5521 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     5581 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5641 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5701 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5761 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5821 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5881 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5941 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6001 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6061 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6121 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6181 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6241 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6301 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6361 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6421 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6481 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6541 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6601 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
     6661 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6721 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6781 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6841 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6901 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6961 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7021 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7081 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7141 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7201 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7261 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7321 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7381 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7441 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7501 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7561 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7621 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     7681 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     7741 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     7801 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     7861 acgaaacacc gggctgcctt gcctctctct gttttagagc tagaaatagc aagttaaaat
     7921 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
     7981 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
     8041 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
     8101 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
     8161 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     8221 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
     8281 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     8341 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     8401 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     8461 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     8521 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     8581 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     8641 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     8701 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     8761 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta
     8821 cctggagcac ctgcctgaaa tcactttttt tcaggttgg
//
