LOCUS       sequence-324514-       13927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of dSaCas9-KRAB and a sgRNA with puromycin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13927)
  AUTHORS   Black JB, McCutcheon SR, Dube S, Barrera A, Klann TS, Rice GA, Adkar
            SS, Soderling SH, Reddy TE, Gersbach CA
  TITLE     Master Regulators and Cofactors of Human Neuronal Cell Fate 
            Specification Identified by CRISPR Gene Activation Screens.
  JOURNAL   Cell Rep. 2020 Dec 1;33(9):108460. doi: 
            10.1016/j.celrep.2020.108460.
  PUBMED    33264623
REFERENCE   2  (bases 1 to 13927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2020-12-1"; volume: "33(9):108460. doi"; pages: " 
            10.1016/j.celrep.2020.108460"
COMMENT     Sequence Label: pLV_hU6-sgRNA_hUbC-dSaCas9-KRAB-T2A-PuroR
FEATURES             Location/Qualifiers
     source          1..13927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        29..133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(134..202,203..994)
                     /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(352..371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1165..1753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1654..1673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1907..1924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2041..2062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2077..2082,2083..2100,2101..2107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2115..2131
                     /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     2120..2142
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2139..2155
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2139..2155
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2204..2325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2215..2234)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2269..2288
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2455..2829)
                     /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"
     promoter        complement(2848..2895)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2943..3272)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2957..3092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3011..3030)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3257..3277
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3286..3714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3396..3417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3608..3627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3760..3984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3973..3990
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             4016..4196
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     4701..4717
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4702..4718
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4721..5309
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4827..4838
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5236..5256
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    5334..5367
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5421..6020)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5504..5524)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6001..6020
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6021..6074)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(6081..6101)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6168..6362)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             complement(6396..6443)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(9624..9644)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9651..9677)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(9715..10926)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(9859..9878)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     11119..11138
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11213..11288
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     misc_feature    11371..11488
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11786..11830)
                     /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    12015..12248
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    12741..12866
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             12913..13093
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(13111..13309)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(13136..13156)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        13311..13690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     13862..13881
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
       61 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      121 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      181 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      241 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      301 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      361 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      421 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      481 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
      541 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
      601 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
      661 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
      721 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
      781 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      841 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      901 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1921 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1981 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2041 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2101 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2161 ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag
     2221 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2281 gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa
     2341 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     2401 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtagca cgtgtcagtc
     2461 ctgctcctcg gccacgaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg
     2521 cccccacggc tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt
     2581 ggacacgacc tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc
     2641 cagggtgttg tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc
     2701 ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt
     2761 ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa
     2821 cttggccatg gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga
     2881 tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg atatacaagc tcccgggagc
     2941 tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag
     3001 gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
     3061 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
     3121 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     3181 ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     3241 cctggggact ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt
     3301 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3361 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3421 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3481 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3541 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3601 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3661 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
     3721 accccctaga gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc
     3781 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     3841 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
     3901 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     3961 atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctgcta
     4021 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     4081 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     4141 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     4201 cagtccggat gcagctctcg ggccatgtga tgaaatgcta ggcggctgtc aaacctccac
     4261 tctaatactt ctctctccgg gtcatccatc ccatgcaggc tcacagggtg taacaagcgg
     4321 gtgttctctc cttcattggc ttcttctacc ttctcttgct caactggtac tagcttgtag
     4381 caccatccaa aggtcagtgg atatctgatc cctggccctg gtgtgtagtt ctgccaatca
     4441 gggaagtagc cttgtgtgtg gtagatccac agatcaagga tatcttgtct tcgttgggag
     4501 tgaattagcc cttccagtcc ccccttttct tttaaaaagt ggctaagatc tacagctgcc
     4561 ttgtaagtca ttggtcttaa aggtacctga ggtgtgactg gaaaacccac ctcctcctcc
     4621 tcttgtgctt ctagccaggc acaatcagca ttggtagctg ctgtattgct acttgtgatt
     4681 gctccatgtt tttctaggtc tcgaggtcga cggtatcgat gcggggaggc ggcccaaagg
     4741 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     4801 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     4861 ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc
     4921 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     4981 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     5041 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     5101 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     5161 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     5221 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     5281 tcacaaattt tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcataactt
     5341 cgtataatgt atgctatacg aagttatcta ttatcaaccg gtcgtgctgc agaagctagt
     5401 aacggccgcc agtgtgctgg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
     5461 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     5521 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     5581 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga cacgccgacg
     5641 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     5701 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     5761 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     5821 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     5881 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     5941 aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc
     6001 gtgggcttgt actcggtcat agggccggga ttctcctcca cgtcaccgca tgttagaaga
     6061 cttcctctgc cctcgctagc aactttgcgt ttctttttcg gaactgatga tttgatttca
     6121 aatgcagtct ctgaatcagg atgggtctct tggtgaattt ctctctccac cagccagggc
     6181 tcttctccct tctccaaccg gaggatcaca tctggcttag taagctgata acccaaggaa
     6241 accaggttct tatagttctc cagcatcaca tttctgtaca ggatctgctg agcagtgtcc
     6301 agcagcttcc actcctccct ggtgaagtcc acaaacacat ccttgaaggt caccagtgtc
     6361 cgggaccagg cagtcagtga cttagcatcg gatccctttt tcttttttgc ctggccggcc
     6421 tttttcgtgg ccgccggcct tttgcccttt ttgatgatct gagggtgctt cttagatttc
     6481 acttcataca ggttgcccag aatgtctgtg ctgtacttct taatgctctg ggtcttggag
     6541 gcgattgtct taatgatcct ggggggcctc ttgtcgttca tgttttccag gtactcgcgg
     6601 taggtgatgt cgatcatgtt cacttcgatc cggttcagca ggtcgttgtt cacgccgatc
     6661 actctataca gctcgccgtt gatcttgatc agatcgttgt tgtagaagga ggcgataaac
     6721 tcggcctggt tgctgatctt cttcagcttc ttagcttcct catagcactt gctattcact
     6781 tcgtagtagt tttctttttt gatcacatcc agattcttca cggtcacgaa cttgtacacg
     6841 ccattgtcca ggtacacgtc gaatctgtag ggcttcaggg acagcttcac gaccttgttt
     6901 ctgctgttgg ggtagtcgtc ggtgatgtcc agatgggcgt tcagtttgtt gccgtaatac
     6961 ttaatcttct tgatcacggg gccgttgtcc tttttggagt acttggtcag gtagttcccg
     7021 gtttcctcgt agtacttgta caggggattc ttctcgtcgc cgtactgttc cataatcagc
     7081 ttcagtttct ggtaggtctg ggggtcgtgg tggtacatca gcagcttttc ggggctcttg
     7141 ttgatcagct ttttcagctt gtcattgtcc ttgtcgtaca ggccgttcag attgttcacg
     7201 atcagggtgt tgcccttgtc gtccttccgg gtggagtaca gggtgtcgtt aatcagctct
     7261 ctattaggct tcttgtccac ccggtggctg tacttgtagt ccttgaagtc cttaatgtgc
     7321 ttgatctggt ggggggtgat gaagatctct ttgtactcct gctcggtttc gatctcgggc
     7381 atgctctcgg cctgcttttc ctcgaacatc tggttttcca tcactttttt ggccttgtcc
     7441 agtttcttcc actctttgaa gatgaaatcg gcgttggcaa tgatcagggc gtcctcggcg
     7501 tggtgcttgt accccttgtt ccgctctttc ttaaacttcc acttccgccg cagaaagctg
     7561 gtgaagccgc cattgatgga cttcactttc acgtccaggt tgttcactct gaagtagctc
     7621 cgcagcaggt tcatcaggcc tctggtggcg tatctggtat ccaccaggtt ccggttgatg
     7681 aagtctttct gcacggagaa cctgttgatg tcccgttctt ccagcagata ctctttcttg
     7741 gtcttgctga ttctgccctt gcccttggcc agattcagga tgtgcttctt gaaggtttcg
     7801 tagctgatct tgctgtcgct gctgctcagg tactggaatg gggtccggtt gcccttcttg
     7861 ctggcttctt cctgcttcac gagcaccttg ttgttgaagc tgttgtcgaa ggacacgctt
     7921 ctggggatga tgtggtccac ctcatagttg aaggggttgt tcagcagatc ttccagaggg
     7981 atggcttcca ggctgtacag gcacttgcct tcctgcatgt cgtgcagctt gatcttctcg
     8041 atcaggtact tggcgttctc tttgccggtg gtccggatga tttcctcgat ccgctcgttg
     8101 gtctgccggt tccgcttctg catctcgttg atcattttct gggcgtcctt ggagttcttc
     8161 tcgcgggcca gctcgataat gatgtcgttg ggcaggccgt acttcttgat gatggcgttg
     8221 atcactttga tgctctggat gaagcttctc ttcacgacgg ggctcaggat gaagtcgtcc
     8281 accagggtgg tggggatctc tttctgctgg gacaggtcca ccttcttggg caccagcttc
     8341 agccggttga agatagcgat ctggttgtcg ttggtgtgcc acagctcgtc caggatcagg
     8401 ttgatggcct tcaggctcag gttgtgggtg ccggtatagc ccttcagatt agagatctgc
     8461 tcgatctctt cctgggtcag ctcggagttc agattggtca gttcttcctg gatgtcctcg
     8521 ctgctctggt agatggtcag gatcttggca atctgatcca gcagctcggc gttctcaata
     8581 atctctttcc gggcggtaat gtccttgatg tcgtggtaca ccttcaggtt ggtgaactcg
     8641 ggcttgccgg tgctggtcac tctgtagccc ttaatatcct cttcgttcac gaggatttct
     8701 ttggcgatct gcttcagggt gggcttcttc ttctgcttga acacgttctc gatgatctgg
     8761 aacttctcgt aatattccag cttctcgttc tcgtccctgg tgatcacgag attgttcagg
     8821 tcgttcaggg cgttgtacag gtcggcgttg taggcgtact tcacgctccg cagttcctcg
     8881 gggaagtagg tgcagtggcc catcagcatc tcgtaccatt ctttgatgtc cttccagccg
     8941 aaggggctgc cctcgccagg tccctcatag taggtccgcc gggtttccag caggtcgatg
     9001 taggtgtcga tgaagctctg gtccagctgg tggtaggcct tctgcacctt cagcagctgt
     9061 ttggcttctt tcacgtagtc gctggtcttg aatctgttga tgctgccccg cacttcgccg
     9121 tctttcttca gccgttccag ctgcagttcg gccacgtatt tctcttccag ggccttgctg
     9181 ttccggctga tctgctcttt ggtggacagc tcgttgccgg tgtcctcttc cacctcgttc
     9241 acgttgtgca cgcctcttct cttggccagg tgcagcaggg cggcagagaa ctcttcctcg
     9301 ctcagcttct ggctcaggcc cttcactctg gcctcgtagg ggttgatgcc gctcagctcg
     9361 ctgtggtcgg tcagcaggtt gtagtcgaac agcagcttct tcactctctg gattctatgc
     9421 cgcctccgcc gcttcagcct tctggcgcct ctcttgctcc gcctaccctc gttgttttcc
     9481 acgttggcct ctttgaacag ccgcacgccg gcatcgatca cgtcccgtgt ctcgtagtcg
     9541 atgatgccgt agcccacgct ggtgatgccg atggccaggc ccaggatgta gttccgcttg
     9601 gctgctggga ctccgtggat accgaccttc cgcttcttct ttggggccat agcgtaatct
     9661 ggaacatcgt atgggtacat ggtggctcta gagtcgacct gcagcccaag cttcgtctaa
     9721 caaaaaagcc aaaaacggcc agaatttagc ggacaattta ctagtctaac actgaaaatt
     9781 acatattgac ccaaatgatt acatttcaaa aggtgcctaa aaaacttcac aaaacacact
     9841 cgccaacccc gagcgcatag ttcaaaaccg gagcttcagc tacttaagaa gataggtaca
     9901 taaaaccgac caaagaaact gacgcctcac ttatccctcc cctcaccaga ggtccggcgc
     9961 ctgtcgattc aggagagcct accctaggcc cgaaccctgc gtcctgcgac ggagaaaagc
    10021 ctaccgcaca cctaccggca ggtggcccca ccctgcatta taagccaaca gaacgggtga
    10081 cgtcacgaca cgacgagggc gcgcgctccc aaaggtacgg gtgcactgcc caacggcacc
    10141 gccataactg ccgcccccgc aacagacgac aaaccgagtt ctccagtcag tgacaaactt
    10201 cacgtcaggg tccccagatg gtgccccagc ccatctcacc cgaataagag ctttcccgca
    10261 ttagcgaagg cctcaagacc ttgggttctt gccgcccacc atgcccccca ccttgtttca
    10321 acgacctcac agcccgcctc acaagcgtct tccattcaag actcgggaac agccgccatt
    10381 ttgctgcgct ccccccaacc cccagttcag ggcaaccttg ctcgcggacc cagactacag
    10441 cccttggcgg tctctccaca cgcttccgtc ccaccgagcg gcccggcggc cacgaaagcc
    10501 ccggccagcc cagcagcccg ctactcacca agtgacgatc acagcgatcc acaaacaaga
    10561 accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc gcgtccttat
    10621 ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg agaagggact
    10681 acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag tcacccaagt
    10741 cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct tatgagcagc
    10801 gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcgctgcgc ccttcgtctg
    10861 acgtggcagc gctcgccgtg aggagggggg cgcccgcggg aggcgccaaa acccggcgcg
    10921 gaggccagat ctggagccga cacgggttaa tgcctatttc ccatgattcc ttcatatttg
    10981 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
    11041 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
    11101 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
    11161 ttggctttat atatcttgtg gaaaggacga aacaccggag acggacgtct ctgttttagt
    11221 actctggaaa cagaatctac taaaacaagg caaaatgccg tgtttatctc gtcaacttgt
    11281 tggcgagatt tttttgcggc cgcccgcggt ggagctccag cttttgttca ttaaccaaac
    11341 tggatctctg ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg
    11401 taattcttta gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta
    11461 ccccccaatc cccccttttc ttttaaaatt gtggatgaat actgccattt gtctgcagaa
    11521 ttggcgcacg cagtgccgat ccgttcacta atcgaatgga tctgtctctg tctctctctc
    11581 caccttcttc ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct
    11641 gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa
    11701 ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata taccacagcc
    11761 aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt
    11821 gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt
    11881 aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag
    11941 atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc
    12001 aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg
    12061 cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag
    12121 ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg
    12181 tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca
    12241 aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg
    12301 gtgctactcc taatggttca atttttacta ctttatattt atataattca cttctccaat
    12361 tgtccctcat atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc
    12421 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
    12481 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
    12541 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
    12601 tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
    12661 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
    12721 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
    12781 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
    12841 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
    12901 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
    12961 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
    13021 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
    13081 accagagaga cccagtacag gcaaaacgcg ctgcttatat agacctccca ccgtacacgc
    13141 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
    13201 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
    13261 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
    13321 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
    13381 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
    13441 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
    13501 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
    13561 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
    13621 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
    13681 aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
    13741 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
    13801 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
    13861 gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc
    13921 cgtcgac
//
