LOCUS EF1a-Cas9-2XNES-2ERT2 14418 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Cas9, 2xNES and 2 tandem ERT2 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS EF1a-Cas9-2XNES-2ERT2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14418)
AUTHORS Zhao C, Zhao Y, Zhang J, Lu J, Chen L, Zhang Y, Ying Y, Xu J, Wei S,
Wang Y
TITLE HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and
Effective Drug Control.
JOURNAL Mol Ther Nucleic Acids. 2018 Dec 7;13:208-219. doi:
10.1016/j.omtn.2018.08.022. Epub 2018 Sep 1.
PUBMED 30312845
REFERENCE 2 (bases 1 to 14418)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14418
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 130..341
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 1205..1214
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1226..5326
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMTKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5465..6400
/codon_start=1
/product="mutated ligand-binding domain of the human
estrogen receptor (Feil et al., 1997)"
/label=ERT2
/translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
CDS 6452..7387
/codon_start=1
/product="mutated ligand-binding domain of the human
estrogen receptor (Feil et al., 1997)"
/label=ERT2
/translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
primer_bind complement(7537..7557)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7542..7871
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7722..7857
/label=SV40 ori
/note="SV40 origin of replication"
CDS 7881..8255
/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"
misc_feature 8341..8929
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8394..8414)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8812..8823)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 9016..9249
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9321..9442
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 9461..9596
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(9616..9635)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9617..9635)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9645..9662)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9645..9661)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9654..9676)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 9803..10258
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9890..9909)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10100..10121
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10284..10388
/gene="bla"
/label=AmpR promoter
CDS 10389..11249
/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(10607..10626)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 11420..12008
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11909..11928
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 12162..12179
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 12296..12317
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 12332..12362
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12370..12386
/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 12375..12397
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 12394..12410
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 12394..12410
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 12429..12449
/label=T3
/note="T3 promoter, forward primer"
promoter 12431..12449
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 12477..12703
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 12704..12884
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 12931..13056
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 13549..13782
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 13967..14011
/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 14278..14395
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 atagcccggg ctcgaccata tgataagctt tgcaaagatg gataaagttt taaacagaga
61 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
121 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
181 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
241 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
301 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
361 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
421 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
481 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
541 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
601 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
661 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
721 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
781 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
841 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
901 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
961 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
1021 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
1081 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
1141 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctcctttc
1201 tagagccacc atgggtggcg cgcctgacaa gaagtacagc atcggcctgg acatcggcac
1261 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
1321 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
1381 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
1441 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tgaccaaggt
1501 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
1561 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
1621 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
1681 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
1741 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
1801 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
1861 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
1921 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
1981 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
2041 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
2101 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
2161 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
2221 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
2281 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
2341 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
2401 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
2461 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
2521 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
2581 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
2641 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
2701 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
2761 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
2821 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
2881 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
2941 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
3001 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
3061 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
3121 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
3181 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
3241 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
3301 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
3361 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
3421 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
3481 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
3541 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
3601 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
3661 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
3721 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
3781 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
3841 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
3901 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
3961 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
4021 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
4081 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
4141 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
4201 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
4261 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
4321 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
4381 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
4441 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
4501 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
4561 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
4621 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
4681 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
4741 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
4801 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
4861 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
4921 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
4981 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
5041 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
5101 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
5161 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
5221 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
5281 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacggag gtagtggagg
5341 tactagttta gccttgaaat tagcaggtct tgatatcgct agcggtacct tagccttgaa
5401 attagcaggt cttgatatca cgcgtgctag ccgcctgctg gaagatggcg atctcgagcc
5461 atctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc
5521 taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga
5581 tgctgagccc cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc
5641 gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc
5701 gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg
5761 tgcctggcta gagatcctga tgattggtct cgtctggcgc tccatggagc acccagtgaa
5821 gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat
5881 ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca
5941 gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt
6001 tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa
6061 gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca
6121 ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg
6181 catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct
6241 ggaggcggcg gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga
6301 ggagacggac caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa
6361 gtattacatc acgggggagg cagagggttt ccctgccaca gctggatccc gcactagtcg
6421 cctgctggaa gatggcgatc tcgagccatc tgctggagac atgagagctg ccaacctttg
6481 gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt ccctgacggc
6541 cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt ccgagtatga
6601 tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc tggcagacag
6661 ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg atttgaccct
6721 ccatgatcag gtccaccttc tagaatgtgc ctggctagag atcctgatga ttggtctcgt
6781 ctggcgctcc atggagcacc cagtgaagct actgtttgct cctaacttgc tcttggacag
6841 gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc tggctacatc
6901 atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca aatctattat
6961 tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc tggaagagaa
7021 ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc tgatggccaa
7081 ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc tcatcctctc
7141 ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga agtgcaagaa
7201 cgtggtgccc ctctatgacc tgctgctgga ggcggcggac gcccaccgcc tacatgcgcc
7261 cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg ccactgcggg
7321 ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag agggtttccc
7381 tgccacagct tgattcgaag tcgacttgtt tattgcagct tataatggtt acaaataaag
7441 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
7501 gtccaaactc atcaatgtat cttatgtcga ccctgtggaa tgtgtgtcag ttagggtgtg
7561 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
7621 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
7681 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
7741 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
7801 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
7861 gcttttgcaa aaagcttacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
7921 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
7981 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
8041 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
8101 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
8161 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
8221 actgcgtgca cttcgtggcc gaggagcagg actgaccgat cgataaaata aaagatttta
8281 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctcgac
8341 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
8401 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
8461 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
8521 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
8581 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
8641 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
8701 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
8761 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
8821 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
8881 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
8941 gctcagtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
9001 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
9061 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
9121 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
9181 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
9241 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
9301 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
9361 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
9421 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
9481 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
9541 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
9601 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
9661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
9721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
9781 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
9841 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
9901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
9961 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
10021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
10081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
10141 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
10201 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
10261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
10321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
10381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
10441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
10501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
10561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
10621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
10681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
10741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
10801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
10861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
10921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
10981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
11041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
11101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
11161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
11221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
11281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
11341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
11401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
11461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
11521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
11581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
11641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
11701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
11761 ccagcttgga gcgaacgacc tacaccaaac tgagatacct acagcgtgag ctatgagaaa
11821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
11881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
11941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
12001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
12061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
12121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
12181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
12241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
12301 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
12361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
12421 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
12481 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
12541 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
12601 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
12661 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
12721 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
12781 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
12841 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
12901 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
12961 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
13021 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
13081 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
13141 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
13201 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
13261 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
13321 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
13381 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
13441 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
13501 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
13561 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
13621 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
13681 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
13741 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
13801 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
13861 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
13921 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
13981 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
14041 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
14101 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
14161 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
14221 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc ggttaacttt
14281 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
14341 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
14401 tcacgagact agcctcga
//
