LOCUS       sequence-237490-       13009 bp    DNA     circular SYN 15-APR-2025
DEFINITION  U6-sgRNA EFS-ProCas9-FlaviS6-P2A-Puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13009)
  AUTHORS   Oakes BL, Fellmann C, Rishi H, Taylor KL, Ren SM, Nadler DC, Yokoo 
            R, Arkin AP, Doudna JA, Savage DF
  TITLE     CRISPR-Cas9 Circular Permutants as Programmable Scaffolds for Genome
            Modification.
  JOURNAL   Cell. 2019 Jan 10;176(1-2):254-267.e16. doi: 
            10.1016/j.cell.2018.11.052.
  PUBMED    30633905
REFERENCE   2  (bases 1 to 13009)
  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: "Cell."; 
            date: "2019-01-10"; volume: "176(1-2):254-267.e16. doi"; pages: " 
            10.1016/j.cell.2018.11.052"
COMMENT     Sequence Label: pCF711
FEATURES             Location/Qualifiers
     source          1..13009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3031..3242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3262..3282
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4300..6801
                     /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
                     IHQSITGLYETRIDLKQKKRGG"
     CDS             7402..7422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7432..7488
                     /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             7489..8085
                     /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     7982..8002
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8101..8689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8154..8174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8572..8583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8761..8994
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9020..9037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9026..9250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9296..9724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9383..9402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9593..9614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9733..9753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9738..10067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9918..10053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9980..9999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10115..10162
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10181..10555
                     /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    10685..10806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10722..10741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10776..10795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10855..10871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10855..10871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10868..10890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10879..10895
                     /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(10903..10909,10910..10927,10928..10933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10948..10969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11086..11103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11257..11845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11337..11356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12016..12807,12808..12876))
                     /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     12639..12658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12877..12981)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggaggacg acgaacgtct
     2881 ctgtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
     2941 acttgaaaaa gtggcaccga gtcggtgctt tttttgaatt cgctagctag gtcttgaaag
     3001 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     3061 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
     3121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     3241 aggaccggtg ccaccatggc cccaaagaag aagcggaagg tcagcgctaa ccccatcaac
     3301 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     3361 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     3421 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     3481 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     3541 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     3601 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     3661 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     3721 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     3781 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     3841 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     3901 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     3961 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     4021 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     4081 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     4141 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     4201 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     4261 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     4321 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     4381 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     4441 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     4501 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     4561 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     4621 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     4681 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     4741 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     4801 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     4861 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     4921 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     4981 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     5041 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     5101 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     5161 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     5221 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     5281 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     5341 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     5401 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     5461 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     5521 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     5581 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     5641 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     5701 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     5761 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     5821 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     5881 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     5941 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     6001 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     6061 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     6121 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     6181 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     6241 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     6301 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     6361 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     6421 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     6481 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     6541 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     6601 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     6661 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     6721 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     6781 aaacaaaaga aacgcggagg caaggacaag aagtacagca tcggcctgga catcggcacc
     6841 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     6901 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     6961 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     7021 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     7081 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     7141 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     7201 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     7261 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     7321 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     7381 cagctgttcg aggaaactag tccaaaaaag aagagaaagg taggatccgg cgcaacaaac
     7441 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag
     7501 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7561 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7621 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7681 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7741 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7801 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7861 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7921 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     7981 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     8041 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     8101 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     8161 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     8221 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8281 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8341 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8401 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8461 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     8521 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8581 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8641 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     8701 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     8761 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     8821 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8881 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     8941 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     9001 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     9061 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     9121 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     9181 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     9241 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     9301 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9361 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9421 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9481 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9541 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9601 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9661 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9721 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     9781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     9841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     9901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     9961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
    10021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
    10081 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
    10141 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
    10201 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
    10261 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
    10321 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
    10381 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    10441 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    10501 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    10561 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    10621 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    10681 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    10741 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    10801 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    10861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    10921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    10981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    11041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    11101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    11161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    11221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    11281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    11341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    11401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    11461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    11521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    11581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    11641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    11701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    11761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    11821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    11881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    11941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    12001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    12061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    12121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    12181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    12241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    12301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    12361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    12421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    12481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    12541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    12601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    12661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    12721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    12781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    12841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    12901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    12961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
