LOCUS       sequence-353139-        8805 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Nuclear Protein Barcode (nPC) for vector/cell tracking. Contains U6 
            trRNA cassette for sgRNA cloning. Enables spatial detection of 
            CRISPR screens..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8805)
  AUTHORS   Dhainaut M, Rose SA, Akturk G, Wroblewska A, Nielsen SR, Park ES, 
            Buckup M, Roudko V, Pia L, Sweeney R, Le Berichel J, Wilk CM, 
            Bektesevic A, Lee BH, Bhardwaj N, Rahman AH, Baccarini A, Gnjatic S,
            Pe'er D, Merad M, Brown BD
  TITLE     Spatial CRISPR genomics identifies regulators of the tumor 
            microenvironment.
  JOURNAL   Cell. 2022 Mar 10. pii: S0092-8674(22)00195-7. doi: 
            10.1016/j.cell.2022.02.015.
  PUBMED    35290801
REFERENCE   2  (bases 1 to 8805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2022 
            Mar 10. pii: S0092-8674(22)00195-7. doi: 
            10.1016/j.cell.2022.02.015."
COMMENT     Sequence Label: pLKO_HSV.Ollas.V5_mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..8805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        139..243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(244..312,313..1104)
                     /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(462..481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1275..1863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1764..1783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2017..2034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2151..2172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2187..2192,2193..2210,2211..2217)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2225..2241
                     /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     2230..2252
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2249..2265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2249..2265
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2284..2304
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2286..2304
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2332..2558
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2559..2739
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2786..2911
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3404..3637
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3822..3866
                     /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"
     promoter        4164..4412
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4164..4184
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4335..4354
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4431..4506
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4554..4671
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4811..5989
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5042..5980
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5937..5957
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6038..6047
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6050..6082
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             6101..6142
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             6164..6205
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6215..6922
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6360..6378)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6628..6647
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6992..7580
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7045..7065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7463..7474)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7655..7888
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7960..8081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7997..8016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8051..8070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8121..8256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8276..8295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8277..8295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8305..8322)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8305..8321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8314..8336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(8463..8805,1..113)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8550..8569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8760..8781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
       61 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
      121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      241 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      301 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      361 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      421 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      481 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      541 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      601 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      661 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      721 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      781 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      841 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
      901 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
      961 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1021 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1081 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1141 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1201 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1261 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1321 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1381 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1441 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1501 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1561 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1621 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1681 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1741 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1801 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1861 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1921 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1981 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2041 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2101 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2161 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2221 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     2281 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     2341 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     2401 gagagaaaaa gcaccgtgca tggcgattgg tggaagtaag gtggtacgat cgtgccttat
     2461 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     2521 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     2581 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     2641 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     2701 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     2761 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     2821 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     2881 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     2941 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     3001 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     3061 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     3121 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     3181 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     3241 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     3301 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     3361 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     3421 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     3481 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     3541 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     3601 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     3661 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     3721 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     3781 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     3841 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     3901 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     3961 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     4021 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     4081 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagt
     4141 atcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     4201 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     4261 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     4321 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     4381 ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct gttttagagc
     4441 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     4501 cggtgctttt tttaattctc gacctcgaga caaatggcag tattcatcca caattttaaa
     4561 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     4621 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     4681 tacagggaca gcagagatcc actttggccg cggctcgagg cgataagctt tgcaaagatg
     4741 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
     4801 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     4861 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     4921 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     4981 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     5041 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     5101 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     5161 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     5221 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     5281 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     5341 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     5401 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     5461 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     5521 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     5581 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     5641 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     5701 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     5761 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     5821 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     5881 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     5941 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattctgca
     6001 gtcgatcgac ggtaccgcgg gcccgggatc cacggtcgcc accatgggcc agcccgagct
     6061 ggcccccgag gaccccgagg acggaggagg cgggggaggg agcggcttcg ccaacgagct
     6121 gggccccagg ctgatgggca agtgcggagg aggcggggga gggggcaaac caattccaaa
     6181 cccactgctg ggactggatt ccacaggcat gctgatggtg agcaagggcg aggaggataa
     6241 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     6301 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     6361 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     6421 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     6481 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     6541 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     6601 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     6661 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     6721 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     6781 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     6841 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     6901 cggcatggac gagctgtaca agaaacggcc agccgaggat atggaggaag aacaagcctt
     6961 taaacgatct cgataaagcg gccgcgtcga caatcaacct ctggattaca aaatttgtga
     7021 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     7081 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     7141 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     7201 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     7261 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     7321 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     7381 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     7441 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     7501 gctgccggct ctgcggcctc ttccgcatct tcgccttcgc cctcagacga gtcggatctc
     7561 cctttgggcc gcctccccgc ctgggtacct ttaagaccaa tgacttacaa ggcagctgta
     7621 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7681 agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7741 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7801 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7861 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     7921 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     7981 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8041 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     8101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     8161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     8221 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     8281 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     8341 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     8401 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     8461 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     8521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     8761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggt
//
