LOCUS       sequence-361347-       12704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral SpCas9-gRNA expression vector with eGFP-P2A-BlastR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12704)
  AUTHORS   Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, 
            Crawford GE, Reddy TE, Gersbach CA
  TITLE     CRISPR-Cas9 epigenome editing enables high-throughput screening for 
            functional regulatory elements in the human genome.
  JOURNAL   Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853.
  PUBMED    28369033
REFERENCE   2  (bases 1 to 12704)
  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: "Nat 
            Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853."
COMMENT     Sequence Label: pLV-U6-gRNA-UbC-eGFP-P2A-Bsr
FEATURES             Location/Qualifiers
     source          1..12704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     378..398
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             441..621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    668..793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1286..1519
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1704..1748
                     /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    2046..2163
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2207..2240)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_RNA        complement(2251..2326)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4208..4456)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4266..4285)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4436..4456)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        4489..5700
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5537..5556
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5729..5731,5732..5734,5735..6445)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5729..5731,5732..5734,5735..6445)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5774..5795)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6035..6054)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6382..6403
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6455..6511
                     /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"
     protein_bind    complement(6944..6977)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7002..7590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7055..7075)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7473..7484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7593..7609)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7594..7610)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8115..8295
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8321..8338)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8327..8551
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8597..9025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8684..8703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8894..8915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9034..9054)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9039..9368
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9219..9354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9281..9300
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9416..9463
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9482..9856
                     /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    9986..10107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10023..10042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10077..10096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10156..10172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10156..10172)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10169..10191)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10180..10196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10204..10210,10211..10228,10229..10234))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10249..10270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10387..10404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10558..11146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10638..10657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11317..12108,12109..12177))
                     /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     11940..11959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12178..12282)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12357..12376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(12548..12704,1..223)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
       61 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      121 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      181 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      241 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      301 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      361 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      421 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      481 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      541 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      601 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      661 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      721 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      781 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      841 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      901 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
      961 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1021 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1081 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1141 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1201 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1261 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1321 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1381 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1441 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1501 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1561 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1621 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1681 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1741 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1801 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1861 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1921 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     1981 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     2041 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2101 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2161 tttcgggttt attacaggga cagcagagat ccagtttggt taattaataa cttcgtatag
     2221 catacattat acgaagttat taaaaaaaaa gcaccgactc ggtgccactt tttcaagttg
     2281 ataacggact agccttattt taacttgcta tttctagctc taaaacagag acgtacaaaa
     2341 aagagcaaga agctaaaaaa gatttaaaaa ttatttttag cgcagttaat ggaacaggaa
     2401 ctaaatttac cccaaaaata ttacgtgaat caggatataa cgttattgag gttgaagagc
     2461 atgcatttga agatgaaaca tttaaaaatg ttgtaaatcc aaatccagaa tttgatcctg
     2521 catgaaaaat accgcttgaa tatggtatta aacatgatgc agatattatt attatgaatg
     2581 acccagatgc tgacagattt ggaatggcaa taaaacatga tggtcatttt gtaagattag
     2641 atggaaatca aacaggacca attttaattg attgaaaatt atcaaatcta aaacgcttaa
     2701 atagcattcc aaaaaatccg gctctatatt caagttttgt aacaagtgat ttgggtgata
     2761 gaatcgctca tgaaaaatat ggagttaata ttgtaaaaac tttaactgga tttaaatgaa
     2821 tgggtagaga aattgctaaa gaagaagata acggattaaa ttttgttttt gcttatgaag
     2881 aaagttatgg atatgtaatt gatgactcag ctagagataa agatggaata caagcttcta
     2941 tattaatagc agaggctgct tgattttata aaaaacaaaa taaaacatta gtagactatt
     3001 tagaagattt atttaaagaa atgggtgcat attacacttt cactttaaac ttgaatttta
     3061 aaccagaaga aaagaaatta aaaattgaac cattaatgaa atcattgaga gcaacaccct
     3121 taactcaaat tgctggactt aaagttgtta atgttgaaga ctacatcgat ggaatgtata
     3181 atatgccagg acaagactta ctaaaatttt atttagaaga taagtcatga tttgctgttc
     3241 gcccaagtgg aactgaacct aaactaaaaa tttattttat aggtgttggt gaatctgttc
     3301 aaaacgctaa agttaaagta gacgaaatta ttaaagaatt aaaattaaaa atgaatatat
     3361 aggagaaaaa atgaaactaa acaaatatat agatcacaca ttattaaaac aagatgctac
     3421 gaaagctgaa attaaacaat tatgtgatga agcaattgaa tttgattttg caacagtttg
     3481 tgttaattca tattgaacaa gctattgtaa agaattatta aaaggcacaa atgtaggaat
     3541 aacaaatgtt gtaggttttc ctctaggtgc atgcacaaca gctacaaaag cattcgaagt
     3601 ttctgaagca attaaagatg gtgcaacaga aattgatatg gtattaaata ttggtgcatt
     3661 aaaagacaaa aattatgaat tagttttaga agacatgaaa gctgtaaaaa aagcagctgg
     3721 atcacatgtt gttaaatgta ttatggaaaa ttgtttatta acaaaagaag aaatcatgaa
     3781 agcttgtgaa atagctgttg aagctggatt agaatttgtt aaaacatcaa caggattttc
     3841 aaaatcaggt gcaacatttg aagatgttaa actaatgaag tcagttgtta aagacaatgc
     3901 tttagttaaa gcagctggtg gagttagaac atttgaagat gctcaaaaaa tgattgaagc
     3961 aggagctgac cgcttaggaa caagtggtgg agtagctatt attaaaggtg aagaaaacaa
     4021 cgcgagttac taaaactagc gtttttttat tttgctcatt tttattaaaa gtttgcaaaa
     4081 aggaacataa aaattctaat tattgatact aaagttatta aaaagaagat tttggttgat
     4141 tttataaagg tcatagaata taatatttta gcatgtgtat tttgtgtgct catttacaac
     4201 cgtctccggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc
     4261 aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc
     4321 caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca
     4381 aattaattcc aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca
     4441 tgggaaatag gccctcttaa ttaattaacc cgtgtcggct ccagatctgg cctccgcgcc
     4501 gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg
     4561 aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     4621 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     4681 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     4741 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     4801 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     4861 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     4921 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     4981 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     5041 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     5101 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg
     5161 aaagctctta ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt
     5221 gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc
     5281 gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc
     5341 gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt
     5401 ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg
     5461 gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc
     5521 tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg
     5581 ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt
     5641 cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac
     5701 gaagcttggg ctgcaggtcg actctagaat ggtgagcaag ggcgaggagc tgttcaccgg
     5761 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     5821 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     5881 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     5941 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     6001 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     6061 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     6121 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     6181 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     6241 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     6301 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     6361 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     6421 tctcggcatg gacgagctgt acaagggatc cggcgcaaca aacttctctc tgctgaaaca
     6481 agccggagat gtcgaagaga atcctggacc gaagaccttc aacatctctc agcaggatct
     6541 ggagctggtg gaggtcgcca ctgagaagat caccatgctc tatgaggaca acaagcacca
     6601 tgtcggggcg gccatcagga ccaagactgg ggagatcatc tctgctgtcc acattgaggc
     6661 ctacattggc agggtcactg tctgtgctga agccattgcc attgggtctg ctgtgagcaa
     6721 cgggcagaag gactttgaca ccattgtggc tgtcaggcac ccctactctg atgaggtgga
     6781 cagatccatc agggtggtca gcccctgtgg catgtgcaga gagctcatct ctgactatgc
     6841 tcctgactgc tttgtgctca ttgagatgaa tggcaagctg gtcaaaacca ccattgagga
     6901 actcatcccc ctcaagtaca ccaggaacta aagttgataa tagataactt cgtatagcat
     6961 acattatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     7021 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     7081 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     7141 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     7201 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     7261 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     7321 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     7381 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     7441 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     7501 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     7561 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa
     7621 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     7681 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     7741 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7801 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     7861 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     7921 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     7981 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     8041 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     8101 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     8161 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     8221 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     8281 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     8341 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     8401 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     8461 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     8521 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     8581 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     8641 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     8701 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8761 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8821 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8881 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     8941 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9001 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     9061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9121 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9181 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     9241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     9301 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     9361 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     9421 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     9481 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     9541 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     9601 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     9661 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     9721 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     9781 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     9841 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     9901 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     9961 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
    10021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    10081 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
    10141 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    10201 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    10261 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    10321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    10381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    10441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    10501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    10561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    10621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    10681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    10741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    10801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    10861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    10921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    10981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    11041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    11101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    11161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    11221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    11281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    11341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    11401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    11461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    11521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    11581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    11641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    11701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    11761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    11821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    11881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    11941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    12001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    12061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    12121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    12181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    12241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    12301 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
    12361 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
    12421 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
    12481 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
    12541 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
    12601 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
    12661 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgt
//
