LOCUS       sequence-160355-       12661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral SpCas9-gRNA expression vector with 
            DsRed-Express2-P2A-BlastR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12661)
  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 12661)
  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-DsRed-P2A-Bsr
FEATURES             Location/Qualifiers
     source          1..12661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             709..889
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    936..1061
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1554..1787
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1972..2016
                     /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    2314..2431
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2475..2508)
                     /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(2519..2594)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4476..4724)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4534..4553)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4704..4724)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        4757..5967
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5804..5823
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5996..6670
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(6176..6196)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     6394..6413
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6588..6611
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             6680..6736
                     /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(7169..7202)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7227..7815
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7280..7300)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7698..7709)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7818..7834)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7819..7835)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8340..8520
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8546..8563)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8552..8776
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8822..9250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8909..8928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9119..9140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9259..9279)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9264..9593
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9444..9579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9506..9525
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9641..9688
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9707..10081
                     /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    10211..10332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10248..10267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10302..10321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10381..10397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10381..10397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10394..10416)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10405..10421
                     /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(10429..10435,10436..10453,10454..10459))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10474..10495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10612..10629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10783..11371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10863..10882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11542..12333,12334..12402))
                     /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     12165..12184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12403..12507)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12582..12601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
       61 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      721 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      781 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      841 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      901 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      961 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1021 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1081 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1141 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1201 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1261 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1321 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1381 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1441 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1501 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1561 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1621 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1681 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1741 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1801 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1861 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1921 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1981 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2041 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2101 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2161 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2221 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     2281 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     2341 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2401 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     2461 agtttggtta attaataact tcgtatagca tacattatac gaagttatta aaaaaaaagc
     2521 accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt
     2581 tctagctcta aaacagagac gtacaaaaaa gagcaagaag ctaaaaaaga tttaaaaatt
     2641 atttttagcg cagttaatgg aacaggaact aaatttaccc caaaaatatt acgtgaatca
     2701 ggatataacg ttattgaggt tgaagagcat gcatttgaag atgaaacatt taaaaatgtt
     2761 gtaaatccaa atccagaatt tgatcctgca tgaaaaatac cgcttgaata tggtattaaa
     2821 catgatgcag atattattat tatgaatgac ccagatgctg acagatttgg aatggcaata
     2881 aaacatgatg gtcattttgt aagattagat ggaaatcaaa caggaccaat tttaattgat
     2941 tgaaaattat caaatctaaa acgcttaaat agcattccaa aaaatccggc tctatattca
     3001 agttttgtaa caagtgattt gggtgataga atcgctcatg aaaaatatgg agttaatatt
     3061 gtaaaaactt taactggatt taaatgaatg ggtagagaaa ttgctaaaga agaagataac
     3121 ggattaaatt ttgtttttgc ttatgaagaa agttatggat atgtaattga tgactcagct
     3181 agagataaag atggaataca agcttctata ttaatagcag aggctgcttg attttataaa
     3241 aaacaaaata aaacattagt agactattta gaagatttat ttaaagaaat gggtgcatat
     3301 tacactttca ctttaaactt gaattttaaa ccagaagaaa agaaattaaa aattgaacca
     3361 ttaatgaaat cattgagagc aacaccctta actcaaattg ctggacttaa agttgttaat
     3421 gttgaagact acatcgatgg aatgtataat atgccaggac aagacttact aaaattttat
     3481 ttagaagata agtcatgatt tgctgttcgc ccaagtggaa ctgaacctaa actaaaaatt
     3541 tattttatag gtgttggtga atctgttcaa aacgctaaag ttaaagtaga cgaaattatt
     3601 aaagaattaa aattaaaaat gaatatatag gagaaaaaat gaaactaaac aaatatatag
     3661 atcacacatt attaaaacaa gatgctacga aagctgaaat taaacaatta tgtgatgaag
     3721 caattgaatt tgattttgca acagtttgtg ttaattcata ttgaacaagc tattgtaaag
     3781 aattattaaa aggcacaaat gtaggaataa caaatgttgt aggttttcct ctaggtgcat
     3841 gcacaacagc tacaaaagca ttcgaagttt ctgaagcaat taaagatggt gcaacagaaa
     3901 ttgatatggt attaaatatt ggtgcattaa aagacaaaaa ttatgaatta gttttagaag
     3961 acatgaaagc tgtaaaaaaa gcagctggat cacatgttgt taaatgtatt atggaaaatt
     4021 gtttattaac aaaagaagaa atcatgaaag cttgtgaaat agctgttgaa gctggattag
     4081 aatttgttaa aacatcaaca ggattttcaa aatcaggtgc aacatttgaa gatgttaaac
     4141 taatgaagtc agttgttaaa gacaatgctt tagttaaagc agctggtgga gttagaacat
     4201 ttgaagatgc tcaaaaaatg attgaagcag gagctgaccg cttaggaaca agtggtggag
     4261 tagctattat taaaggtgaa gaaaacaacg cgagttacta aaactagcgt ttttttattt
     4321 tgctcatttt tattaaaagt ttgcaaaaag gaacataaaa attctaatta ttgatactaa
     4381 agttattaaa aagaagattt tggttgattt tataaaggtc atagaatata atattttagc
     4441 atgtgtattt tgtgtgctca tttacaaccg tctccggtgt ttcgtccttt ccacaagata
     4501 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
     4561 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
     4621 tgtactaata tctttgtgtt tacagtcaaa ttaattccaa ttatctctct aacagccttg
     4681 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttaatt aattaacccg
     4741 tgtcggctcc agatctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     4801 ctcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgttcctga tccttccgcc
     4861 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     4921 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     4981 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     5041 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     5101 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     5161 tgcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     5221 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     5281 ggggttgggg ggagcgcaca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     5341 taaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     5401 aggtcttgag gccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca
     5461 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcgggt ttgtcgtctg
     5521 gttgcggggg cggcagttat gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     5581 gcgcctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     5641 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     5701 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     5761 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     5821 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     5881 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     5941 tggccgtttt tggctttttt gttagacgaa gcttgggctg caggtcgact ctagaatgga
     6001 tagcactgag aacgtcatca agcccttcat gcgcttcaag gtgcacatgg agggctccgt
     6061 gaacggccac gagttcgaga tcgagggcga gggcgagggc aagccctacg agggcaccca
     6121 gaccgccaag ctgcaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     6181 cccccagttc cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta
     6241 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     6301 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcaccttca tctaccacgt
     6361 gaagttcatc ggcgtgaact tcccctccga cggccccgta atgcagaaga agactctggg
     6421 ctgggagccc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     6481 caaggcgctg aagctgaagg gcggcggcca ctacctggtg gagttcaagt caatctacat
     6541 ggccaagaag cccgtgaagc tgcccggcta ctactacgtg gactccaagc tggacatcac
     6601 ctcccacaac gaggactaca ccgtggtgga gcagtacgag cgcgccgagg cccgccacca
     6661 cctgttccag ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga
     6721 agagaatcct ggaccgaaga ccttcaacat ctctcagcag gatctggagc tggtggaggt
     6781 cgccactgag aagatcacca tgctctatga ggacaacaag caccatgtcg gggcggccat
     6841 caggaccaag actggggaga tcatctctgc tgtccacatt gaggcctaca ttggcagggt
     6901 cactgtctgt gctgaagcca ttgccattgg gtctgctgtg agcaacgggc agaaggactt
     6961 tgacaccatt gtggctgtca ggcaccccta ctctgatgag gtggacagat ccatcagggt
     7021 ggtcagcccc tgtggcatgt gcagagagct catctctgac tatgctcctg actgctttgt
     7081 gctcattgag atgaatggca agctggtcaa aaccaccatt gaggaactca tccccctcaa
     7141 gtacaccagg aactaaagtt gataatagat aacttcgtat agcatacatt atacgaagtt
     7201 atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     7261 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     7321 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     7381 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     7441 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     7501 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     7561 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     7621 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     7681 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     7741 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     7801 gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca
     7861 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     7921 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     7981 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     8041 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     8101 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     8161 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     8221 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     8281 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
     8341 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     8401 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     8461 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     8521 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     8581 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     8641 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     8701 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     8761 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     8821 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     8881 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     8941 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     9001 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     9061 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     9121 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     9181 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     9241 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     9301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     9361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     9421 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     9481 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     9541 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     9601 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     9661 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     9721 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     9781 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     9841 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     9901 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     9961 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
    10021 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
    10081 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
    10141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
    10201 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    10261 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    10321 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
    10381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    10441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    10501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    10561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    10621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    10681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    10741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    10801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    10861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    10921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    10981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    11041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    11101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    11161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    11221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    11281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    11341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    11401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    11461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    11521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    11581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    11641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    11701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    11761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    11821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    11881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    11941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    12001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    12061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    12121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    12181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    12241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    12301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    12361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    12421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    12481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
    12541 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
    12601 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
    12661 a
//
