LOCUS       sequence-353011-        8772 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 8772)
  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 8772)
  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_HA.HSV.NWS_mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..8772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     652..672
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        654..672
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        700..926
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             927..1107
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1154..1279
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1772..2005
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2190..2234
                     /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        2532..2780
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2532..2552
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2703..2722
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2799..2874
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2922..3039
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3179..4357
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3410..4348
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4305..4325
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4406..4415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4418..4444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4463..4495
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             4517..4540
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4550..5257
                     /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(4695..4713)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4963..4982
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5327..5915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5380..5400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5798..5809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5990..6223
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6295..6416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6332..6351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6386..6405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6456..6591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6611..6630)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6612..6630)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6640..6657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6640..6656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6649..6671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6798..7253
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6885..6904)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7095..7116
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7279..7383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7384..7452,7453..8244)
                     /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(7602..7621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8415..8772,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      661 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      721 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      781 accgtgcatg gcgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      841 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      901 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      961 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1021 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1081 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     1141 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     1201 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     1261 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1321 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1381 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1441 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1501 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1561 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1621 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1681 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1741 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1801 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1861 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1921 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1981 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     2041 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     2101 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     2161 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2221 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2281 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2341 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2401 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2461 ccattcgatt agtgaacgga tctcgacggt atcgatcacg agactagtat cgagcggccg
     2521 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     2581 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     2641 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     2701 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     2761 tgtggaaagg acgaaacacc gggtcttcga gaagacctgt tttagagcta gaaatagcaa
     2821 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     2881 taattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2941 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     3001 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     3061 agagatccac tttggccgcg gctcgaggcg ataagctttg caaagatgga taaagtttta
     3121 aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg
     3181 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     3241 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     3301 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     3361 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
     3421 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
     3481 ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
     3541 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc
     3601 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
     3661 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
     3721 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
     3781 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
     3841 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
     3901 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
     3961 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
     4021 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
     4081 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
     4141 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
     4201 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
     4261 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
     4321 ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attctgcagt cgatcgacgg
     4381 taccgcgggc ccgggatcca cggtcgccac catgggctac ccatacgatg tgccagatta
     4441 cgctggagga ggcgggggag ggcagcccga gctggccccc gaggaccccg aggacggagg
     4501 aggcggggga gggaactggt cccaccccca attcgagaaa ggcatgctga tggtgagcaa
     4561 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     4621 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     4681 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     4741 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     4801 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     4861 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     4921 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     4981 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     5041 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     5101 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     5161 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     5221 ccgccactcc accggcggca tggacgagct gtacaagaaa cggccagccg aggatatgga
     5281 ggaagaacaa gcctttaaac gatctcgata aagcggccgc gtcgacaatc aacctctgga
     5341 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5401 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5461 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5521 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5581 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5641 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5701 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5761 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5821 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg catcttcgcc ttcgccctca
     5881 gacgagtcgg atctcccttt gggccgcctc cccgcctggg tacctttaag accaatgact
     5941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6001 attcactccc aacgaagata agatctgctt tttgcttgta ctgggtctct ctggttagac
     6061 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6241 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6421 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6481 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6541 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     6601 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     6661 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6721 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6781 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6841 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6901 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6961 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7021 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7081 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7141 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7201 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     7261 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7321 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7381 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7441 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7501 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7561 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7621 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7681 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7741 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7801 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7861 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7921 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7981 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8041 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8101 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8161 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8221 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8281 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8521 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8761 ttggagcgaa cg
//
