LOCUS       sequence-352851-        8793 bp    DNA     circular SYN 15-DEC-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 8793)
  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 8793)
  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. 2022 
            Mar 10. pii: S0092-8674(22)00195-7. doi: 
            10.1016/j.cell.2022.02.015."
COMMENT     Sequence Label: pLKO_C.NWS.Ollas_mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..8793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(220..288,289..1080)
                     /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(438..457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2127..2148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2163..2168,2169..2186,2187..2193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2201..2217
                     /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     2206..2228
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2225..2241
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2225..2241
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2260..2280
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2262..2280
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2308..2534
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2535..2715
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2762..2887
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3380..3613
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3798..3842
                     /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        4140..4388
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4140..4160
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4311..4330
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4407..4482
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4530..4647
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4787..5965
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5018..5956
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5913..5933
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6014..6023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6026..6061
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             6083..6106
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6125..6166
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             6179..6886
                     /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(6324..6342)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6592..6611
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6956..7544
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7009..7029)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7427..7438)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7619..7852
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7924..8045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7961..7980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8015..8034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8085..8220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8240..8259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8241..8259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8269..8286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8269..8285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8278..8300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(8427..8793,1..89)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8514..8533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8724..8745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
       61 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      181 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      241 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      301 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      361 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      421 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
      481 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      541 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      601 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      661 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      721 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
      781 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      841 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      901 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      961 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1021 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1081 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2041 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2101 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2161 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2221 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     2281 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     2341 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatggc
     2401 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     2461 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     2521 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2581 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2641 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2701 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     2761 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     2821 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     2881 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     2941 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     3001 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     3061 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     3121 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     3181 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     3241 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     3301 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     3361 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     3421 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     3481 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     3541 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     3601 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     3661 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     3721 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     3781 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     3841 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     3901 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     3961 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     4021 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     4081 tgaacggatc tcgacggtat cgatcacgag actagtatcg agcggccgcc cccttcaccg
     4141 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     4201 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     4261 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     4321 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     4381 gaaacaccgg gtcttcgaga agacctgttt tagagctaga aatagcaagt taaaataagg
     4441 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttta attctcgacc
     4501 tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     4561 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     4621 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     4681 tggccgcggc tcgaggcgat aagctttgca aagatggata aagttttaaa cagagaggaa
     4741 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     4801 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     4861 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     4921 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     4981 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     5041 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     5101 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     5161 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     5221 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     5281 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     5341 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     5401 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     5461 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     5521 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     5581 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     5641 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     5701 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     5761 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     5821 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     5881 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     5941 ttttcttcca tttcaggtgt cgtgaggaat tctgcagtcg atcgacggta ccgcgggccc
     6001 gggatccacg gtcgccacca tgggcgagga ccaggtggac cccaggctga tcgacggcaa
     6061 gggaggaggc gggggaggga actggtccca cccccaattc gagaaaggag gaggcggggg
     6121 agggagcggc ttcgccaacg agctgggccc caggctgatg ggcaagtgcg gcatgctgat
     6181 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     6241 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     6301 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     6361 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     6421 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     6481 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     6541 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     6601 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     6661 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     6721 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     6781 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     6841 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagaaac ggccagccga
     6901 ggatatggag gaagaacaag cctttaaacg atctcgataa agcggccgcg tcgacaatca
     6961 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     7021 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     7081 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     7141 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     7201 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     7261 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     7321 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     7381 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7441 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc atcttcgcct
     7501 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgggt acctttaaga
     7561 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     7621 gaagggctaa ttcactccca acgaagataa gatctgcttt ttgcttgtac tgggtctctc
     7681 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7741 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7801 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     7861 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     7921 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7981 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8041 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     8101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     8161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     8221 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     8281 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     8341 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     8401 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     8461 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     8521 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     8581 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     8641 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     8701 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     8761 tatctcggtc tattcttttg atttataagg gat
//
