LOCUS       Exported               12073 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral vector encoding modified SaCas9 system backbone bearing 
            BsmBI site for new guide RNAs and puromycin selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    pCK002_U6-Sa-sgRNA(mod)_EFS-SaCas9-2A-Puro_WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12073)
  AUTHORS   Singer M, Wang C, Cong L, Marjanovic ND, Kowalczyk MS, Zhang H, 
            Nyman J, Sakuishi K, Kurtulus S, Gennert D, Xia J, Kwon JY, Nevin J,
            Herbst RH, Yanai I, Rozenblatt-Rosen O, Kuchroo VK, Regev A, 
            Anderson AC
  TITLE     A Distinct Gene Module for Dysfunction Uncoupled from Activation in 
            Tumor-Infiltrating T Cells.
  JOURNAL   Cell. 2016 Sep 8;166(6):1500-1511.e9. doi: 
            10.1016/j.cell.2016.08.052.
  PUBMED    27610572
REFERENCE   2  (bases 1 to 12073)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..2848
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2873..2948
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        3007..3218
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3226..3235
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3238..3258
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3283..6438
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             6439..6486
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6496..6552
                     /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             6553..7149
                     /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     7046..7066
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7165..7753
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7218..7238)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7636..7647)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7825..8058
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8084..8101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8090..8314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8360..8788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8447..8466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8657..8678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8797..8817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8802..9131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8982..9117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9044..9063
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9179..9226
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9245..9619
                     /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    9749..9870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9786..9805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9840..9859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9919..9935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9919..9935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9932..9954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9943..9959
                     /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(9967..9997)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10012..10033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10150..10167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10321..10909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10401..10420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11080..11940)
                     /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     11703..11722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11941..12045)
                     /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 attggaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttacagtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggacgtct ctgttctagt
     2881 actctggaaa cagaatctac tagaacaagg caaaatgccg tgtttatctc gtcaacttgt
     2941 tggcgagatt tttgatcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc
     3001 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3061 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3121 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3181 acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggtgccac catggcccca
     3241 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccaagcggaa ctacatcctg
     3301 ggcctggaca tcggcatcac cagcgtgggc tacggcatca tcgactacga gacacgggac
     3361 gtgatcgatg ccggcgtgcg gctgttcaaa gaggccaacg tggaaaacaa cgagggcagg
     3421 cggagcaaga gaggcgccag aaggctgaag cggcggaggc ggcatagaat ccagagagtg
     3481 aagaagctgc tgttcgacta caacctgctg accgaccaca gcgagctgag cggcatcaac
     3541 ccctacgagg ccagagtgaa gggcctgagc cagaagctga gcgaggaaga gttctctgcc
     3601 gccctgctgc acctggccaa gagaagaggc gtgcacaacg tgaacgaggt ggaagaggac
     3661 accggcaacg agctgtccac caaagagcag atcagccgga acagcaaggc cctggaagag
     3721 aaatacgtgg ccgaactgca gctggaacgg ctgaagaaag acggcgaagt gcggggcagc
     3781 atcaacagat tcaagaccag cgactacgtg aaagaagcca aacagctgct gaaggtgcag
     3841 aaggcctacc accagctgga ccagagcttc atcgacacct acatcgacct gctggaaacc
     3901 cggcggacct actatgaggg acctggcgag ggcagcccct tcggctggaa ggacatcaaa
     3961 gaatggtacg agatgctgat gggccactgc acctacttcc ccgaggaact gcggagcgtg
     4021 aagtacgcct acaacgccga cctgtacaac gccctgaacg acctgaacaa tctcgtgatc
     4081 accagggacg agaacgagaa gctggaatat tacgagaagt tccagatcat cgagaacgtg
     4141 ttcaagcaga agaagaagcc caccctgaag cagatcgcca aagaaatcct cgtgaacgaa
     4201 gaggatatta agggctacag agtgaccagc accggcaagc ccgagttcac caacctgaag
     4261 gtgtaccacg acatcaagga cattaccgcc cggaaagaga ttattgagaa cgccgagctg
     4321 ctggatcaga ttgccaagat cctgaccatc taccagagca gcgaggacat ccaggaagaa
     4381 ctgaccaatc tgaactccga gctgacccag gaagagatcg agcagatctc taatctgaag
     4441 ggctataccg gcacccacaa cctgagcctg aaggccatca acctgatcct ggacgagctg
     4501 tggcacacca acgacaacca gatcgctatc ttcaaccggc tgaagctggt gcccaagaag
     4561 gtggacctgt cccagcagaa agagatcccc accaccctgg tggacgactt catcctgagc
     4621 cccgtcgtga agagaagctt catccagagc atcaaagtga tcaacgccat catcaagaag
     4681 tacggcctgc ccaacgacat cattatcgag ctggcccgcg agaagaactc caaggacgcc
     4741 cagaaaatga tcaacgagat gcagaagcgg aaccggcaga ccaacgagcg gatcgaggaa
     4801 atcatccgga ccaccggcaa agagaacgcc aagtacctga tcgagaagat caagctgcac
     4861 gacatgcagg aaggcaagtg cctgtacagc ctggaagcca tccctctgga agatctgctg
     4921 aacaacccct tcaactatga ggtggaccac atcatcccca gaagcgtgtc cttcgacaac
     4981 agcttcaaca acaaggtgct cgtgaagcag gaagaaaaca gcaagaaggg caaccggacc
     5041 ccattccagt acctgagcag cagcgacagc aagatcagct acgaaacctt caagaagcac
     5101 atcctgaatc tggccaaggg caagggcaga atcagcaaga ccaagaaaga gtatctgctg
     5161 gaagaacggg acatcaacag gttctccgtg cagaaagact tcatcaaccg gaacctggtg
     5221 gataccagat acgccaccag aggcctgatg aacctgctgc ggagctactt cagagtgaac
     5281 aacctggacg tgaaagtgaa gtccatcaat ggcggcttca ccagctttct gcggcggaag
     5341 tggaagttta agaaagagcg gaacaagggg tacaagcacc acgccgagga cgccctgatc
     5401 attgccaacg ccgatttcat cttcaaagag tggaagaaac tggacaaggc caaaaaagtg
     5461 atggaaaacc agatgttcga ggaaaagcag gccgagagca tgcccgagat cgaaaccgag
     5521 caggagtaca aagagatctt catcaccccc caccagatca agcacattaa ggacttcaag
     5581 gactacaagt acagccaccg ggtggacaag aagcctaata gagagctgat taacgacacc
     5641 ctgtactcca cccggaagga cgacaagggc aacaccctga tcgtgaacaa tctgaacggc
     5701 ctgtacgaca aggacaatga caagctgaaa aagctgatca acaagagccc cgaaaagctg
     5761 ctgatgtacc accacgaccc ccagacctac cagaaactga agctgattat ggaacagtac
     5821 ggcgacgaga agaatcccct gtacaagtac tacgaggaaa ccgggaacta cctgaccaag
     5881 tactccaaaa aggacaacgg ccccgtgatc aagaagatta agtattacgg caacaaactg
     5941 aacgcccatc tggacatcac cgacgactac cccaacagca gaaacaaggt cgtgaagctg
     6001 tccctgaagc cctacagatt cgacgtgtac ctggacaatg gcgtgtacaa gttcgtgacc
     6061 gtgaagaatc tggatgtgat caaaaaagaa aactactacg aagtgaatag caagtgctat
     6121 gaggaagcta agaagctgaa gaagatcagc aaccaggccg agtttatcgc ctccttctac
     6181 aacaacgatc tgatcaagat caacggcgag ctgtatagag tgatcggcgt gaacaacgac
     6241 ctgctgaacc ggatcgaagt gaacatgatc gacatcacct accgcgagta cctggaaaac
     6301 atgaacgaca agaggccccc caggatcatt aagacaatcg cctccaagac ccagagcatt
     6361 aagaagtaca gcacagacat tctgggcaac ctgtatgaag tgaaatctaa gaagcaccct
     6421 cagatcatca aaaagggcaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     6481 aaaaagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     6541 aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     6601 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     6661 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     6721 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     6781 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     6841 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     6901 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
     6961 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7021 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7081 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7141 ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     7201 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     7261 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     7321 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     7381 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     7441 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     7501 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     7561 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     7621 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     7681 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     7741 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     7801 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     7861 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7921 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7981 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8041 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     8101 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     8161 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     8221 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     8281 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     8341 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     8461 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8521 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8581 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8641 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8701 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8761 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     8821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     8881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     8941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     9001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     9061 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     9121 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     9181 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     9241 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     9301 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     9361 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     9421 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     9481 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     9541 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     9601 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     9661 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     9721 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     9781 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     9841 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     9901 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     9961 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    10021 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    10081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    10141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    10201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    10261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    10321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    10441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    10501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    10561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    10621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    10681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    10741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    10801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    10861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    10921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    10981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    11041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    11101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    11161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    11221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    11281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    11341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    11401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    11461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    11521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    11581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    11641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    11701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    11761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    11821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    11881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    11941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    12001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    12061 agtgccacct gac
//
