LOCUS       pGMC00006_Lenti-CRISPR-V2_U6-Sp-ASNS-IVT-538       13013 bp ds-DNA     circular SYN 01-FEB-2021
DEFINITION  sgRNA targeting ASNS cloned into Lenti-CRISPR-V2.
ACCESSION
VERSION     .
KEYWORDS    lentiCRISPR v2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 14873)
  AUTHORS   Sanjana, NE, Shalem, O, Zhang and F
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 14873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13013
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..199
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /standard_name="CMV promoter"
     primer_bind     154..174
                     /note="Human CMV immediate early promoter, forward primer"
                     /standard_name="CMV-F"
     LTR             217..397
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /standard_name="5' LTR (truncated)"
     misc_feature    444..569
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /standard_name="HIV-1 Psi"
     misc_feature    1062..1295
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to
                     thecytoplasm."
                     /standard_name="RRE"
     misc_feature    1822..1939
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /standard_name="cPPT/CTS"
     promoter        1990..2230
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     1990..2010
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     2161..2180
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     PCR_primer      2217..2239
                     /standard_name="Primer_592"
     misc_RNA        2240..2259
                     /standard_name="ASNS-IVT-538"
     misc_feature    2242..2259
                     /standard_name="GACGGTTGTAAATGAGCACACAAAATACACATGCTAAAATAT
                     TATATTCTATGACCTTTATAAAATCAACCAAAATCTTCTTTTTAATAACTTTAGTATC
                     AATAATTAGAATTTTTATGTTCCTTTTTGCAAACTTTTAATAAAAATGAGCAAAATAA
                     AAAAACGCTAGTTTTAGTAACTCGCGTTGTTTTCTTCACCTTTAATAATAGCTACTCC
                     ACCACTTGTTCCTAAGCGGTCAGCTCCTGCTTCAATCATTTTTTGAGCATCTTCAAAT
                     GTTCTAACTCCACCAGCTGCTTTAACTAAAGCATTGTCTTTAACAACTGACTTCATTA
                     GTTTAACATCTTCAAATGTTGCACCTGATTTTGAAAATCCTGTTGATGTTTTAACAAA
                     TTCTAATCCAGCTTCAACAGCTATTTCACAAGCTTTCATGATTTCTTCTTTTGTTAAT
                     AAACAATTTTCCATAATACATTTAACAACATGTGATCCAGCTGCTTTTTTTACAGCTT
                     TCATGTCTTCTAAAACTAATTCATAATTTTTGTCTTTTAATGCACCAATATTTAATAC
                     CATATCAATTTCTGTTGCACCATCTTTAATTGCTTCAGAAACTTCGAATGCTTTTGTA
                     GCTGTTGTGCATGCACCTAGAGGAAAACCTACAACATTTGTTATTCCTACATTTGTGC
                     CTTTTAATAATTCTTTACAATAGCTTGTTCAATATGAATTAACACAAACTGTTGCAAA
                     ATCAAATTCAATTGCTTCATCACATAATTGTTTAATTTCAGCTTTCGTAGCATCTTGT
                     TTTAATAATGTGTGATCTATATATTTGTTTAGTTTCATTTTTTCTCCTATATATTCAT
                     TTTTAATTTTAATTCTTTAATAATTTCGTCTACTTTAACTTTAGCGTTTTGAACAGAT
                     TCACCAACACCTATAAAATAAATTTTTAGTTTAGGTTCAGTTCCACTTGGGCGAACAG
                     CAAATCATGACTTATCTTCTAAATAAAATTTTAGTAAGTCTTGTCCTGGCATATTATA
                     CATTCCATCGATGTAGTCTTCAACATTAACAACTTTAAGTCCAGCAATTTGAGTTAAG
                     GGTGTTGCTCTCAATGATTTCATTAATGGTTCAATTTTTAATTTCTTTTCTTCTGGTT
                     TAAAATTCAAGTTTAAAGTGAAAGTGTAATATGCACCCATTTCTTTAAATAAATCTTC
                     TAAATAGTCTACTAATGTTTTATTTTGTTTTTTATAAAATCAAGCAGCCTCTGCTATT
                     AATATAGAAGCTTGTATTCCATCTTTATCTCTAGCTGAGTCATCAATTACATATCCAT
                     AACTTTCTTCATAAGCAAAAACAAAATTTAATCCGTTATCTTCTTCTTTAGCAATTTC
                     TCTACCCATTCATTTAAATCCAGTTAAAGTTTTTACAATATTAACTCCATATTTTTCA
                     TGAGCGATTCTATCACCCAAATCACTTGTTACAAAACTTGAATATAGAGCCGGATTTT
                     TTGGAATGCTATTTAAGCGTTTTAGATTTGATAATTTTCAATCAATTAAAATTGGTCC
                     TGTTTGATTTCCATCTAATCTTACAAAATGACCATCATGTTTTATTGCCATTCCAAAT
                     CTGTCAGCATCTGGGTCATTCATAATAATAATATCTGCATCATGTTTAATACCATATT
                     CAAGCGGTATTTTTCATGCAGGATCAAATTCTGGATTTGGATTTACAACATTTTTAAA
                     TGTTTCATCTTCAAATGCATGCTCTTCAACCTCAATAACGTTATATCCTGATTCACGT
                     AATATTTTTGGGGTAAATTTAGTTCCTGTTCCATTAACTGCGCTAAAAATAATTTTTA
                     AATCTTTTTTAGCTTCTTGCTCTTTTTTGTACGTCTCT"
                     /note="Geneious type: Editing History Replacement"
     misc_RNA        2260..2335
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /standard_name="gRNA scaffold"
     promoter        2397..2608
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
                     /standard_name="EF-1-alpha core promoter"
     PCR_primer      complement(2447..2466)
                     /standard_name="Primer_667"
     regulatory      2627..2636
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong
                     initiationof translation (Kozak, 1987)"
     CDS             2633..6736
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIK
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
                     ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH
                     MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK
                     SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD
                     NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL
                     LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK
                     LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY
                     YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL
                     PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE
                     DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT
                     LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF
                     LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT
                     VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE
                     HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV
                     LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA
                     GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG
                     KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS
                     MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV
                     AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF
                     ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH
                     KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA
                     PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
                     /standard_name="Cas9"
     CDS             6737..6784
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /translation="KRPAATKKAGQAKKKK"
                     /standard_name="nucleoplasmin NLS"
     CDS             6785..6808
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an
                     enterokinasecleavage site"
                     /translation="DYKDDDDK"
                     /standard_name="FLAG"
     CDS             6818..6874
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding
                     separatepolypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /standard_name="P2A"
     CDS             6875..7471
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLE
                     TSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
                     /standard_name="PuroR"
     primer_bind     7368..7388
                     /note="Puromycin resistance gene, forward primer"
                     /standard_name="Puro-F"
     misc_feature    7487..8075
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /standard_name="WPRE"
     primer_bind     complement(7540..7560)
                     /note="WPRE, reverse primer"
                     /standard_name="WPRE-R"
     CDS             complement(7958..7969)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /standard_name="Factor Xa site"
     LTR             8147..8380
                     /note="self-inactivating 3' long terminal repeat (LTR)
                     from HIV-1"
                     /standard_name="3' LTR (Delta-U3)"
     primer_bind     complement(8406..8423)
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /standard_name="BGH-rev"
     polyA_signal    8412..8636
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8682..9110
     primer_bind     complement(8769..8788)
                     /note="F1 origin, reverse primer"
                     /standard_name="F1ori-R"
     primer_bind     8979..9000
                     /note="F1 origin, forward primer"
                     /standard_name="F1ori-F"
     primer_bind     complement(9119..9139)
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /standard_name="pBABE 3'"
     promoter        9124..9453
                     /note="SV40 enhancer and early promoter"
                     /standard_name="SV40 promoter"
     rep_origin      9304..9439
     primer_bind     9366..9385
                     /note="SV40 promoter/origin, forward primer"
                     /standard_name="SV40pro-F"
     promoter        9501..9548
                     /note="synthetic bacterial promoter"
                     /standard_name="EM7 promoter"
     CDS             9567..9941
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW
                     GREFALRDPAGNCVHFVAEEQD"
                     /standard_name="BleoR"
     polyA_signal    10071..10192
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10108..10127)
                     /note="SV40 polyA, reverse primer"
                     /standard_name="SV40pA-R"
     primer_bind     10162..10181
                     /note="SV40 polyA terminator, reverse primer"
                     /standard_name="EBV-rev"
     primer_bind     complement(10241..10257)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /standard_name="M13 rev"
     primer_bind     complement(10241..10257)
                     /note="In lacZ gene. Also called M13-rev"
                     /standard_name="M13 Reverse"
     primer_bind     complement(10254..10276)
                     /note="In lacZ gene"
                     /standard_name="M13/pUC Reverse"
     protein_bind    10265..10281
                     /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)."
                     /standard_name="lac operator"
     promoter        complement(10289..10319)
                     /note="promoter for the E. coli lac operon"
                     /standard_name="lac promoter"
     protein_bind    10334..10355
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
                     /standard_name="CAP binding site"
     primer_bind     complement(10472..10489)
                     /note="L4440 vector, forward primer"
                     /standard_name="L4440"
     rep_origin      complement(10643..11231)
     primer_bind     complement(10723..10742)
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     CDS             complement(11402..12262)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
     primer_bind     12025..12044
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     promoter        complement(12263..12367)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     primer_bind     complement(12442..12461)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /standard_name="pRS-marker"
     enhancer        12633..13012
                     /note="human cytomegalovirus immediate early enhancer"
                     /standard_name="CMV 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 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccgg atcgaactac tgctgcccag ttttagagct agaaatagca
     2281 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     2341 tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     2401 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     2461 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     2521 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     2581 tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca ccatggacaa
     2641 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
     2701 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     2761 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     2821 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     2881 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     2941 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     3001 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     3061 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     3121 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     3181 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     3241 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     3301 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     3361 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     3421 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     3481 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     3541 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     3601 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     3661 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     3721 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     3781 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     3841 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     3901 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     3961 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     4021 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     4081 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     4141 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     4201 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     4261 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     4321 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     4381 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     4441 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     4501 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     4561 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     4621 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     4681 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     4741 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     4801 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     4861 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     4921 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     4981 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     5041 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     5101 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     5161 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     5221 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     5281 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     5341 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     5401 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     5461 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     5521 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     5581 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     5641 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     5701 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     5761 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     5821 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     5881 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     5941 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     6001 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     6061 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     6121 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     6181 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     6241 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     6301 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     6361 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     6421 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     6481 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     6541 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     6601 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     6661 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     6721 tcagctggga ggcgacaagc gacctgccgc cacaaagaag gctggacagg ctaagaagaa
     6781 gaaagattac aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa
     6841 acaagccgga gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct
     6901 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     6961 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     7021 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     7081 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     7141 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     7201 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt
     7261 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     7321 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     7381 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     7441 ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga
     7501 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     7561 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     7621 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     7681 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     7741 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     7801 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     7861 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     7921 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     7981 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     8041 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
     8101 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     8161 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     8221 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     8281 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     8341 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
     8401 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     8461 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     8521 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     8581 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     8641 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     8701 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     8761 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     8821 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     8881 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     8941 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     9001 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     9061 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     9121 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     9181 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     9241 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     9301 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     9361 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     9421 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     9481 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     9541 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     9601 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     9661 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     9721 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     9781 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     9841 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     9901 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     9961 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
    10021 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
    10081 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    10141 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
    10201 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
    10261 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    10321 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    10381 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    10441 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    10501 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    10561 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    10621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    10681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    10741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    10801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    10861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    10921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    10981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    11041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    11101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    11161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    11221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    11281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    11341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    11401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    11461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    11521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    11581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    11641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    11701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    11761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    11821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    11881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    11941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    12001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    12061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    12121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    12181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    12241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    12301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    12361 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
    12421 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
    12481 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
    12541 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
    12601 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
    12661 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
    12721 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
    12781 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
    12841 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
    12901 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
    12961 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgg
//
