LOCUS       sequence-334985-        8835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein Barcode (Pro-Code) for vector/cell tracking. Contains U6 
            trRNA cassette for sgRNA cloning. Enables phenotypic CRISPR screens 
            at a single cell resolution (using cytometry)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8835)
  AUTHORS   Wroblewska A, Dhainaut M, Ben-Zvi B, Rose SA, Park ES, Amir ED, 
            Bektesevic A, Baccarini A, Merad M, Rahman AH, Brown BD
  TITLE     Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens.
  JOURNAL   Cell. 2018 Nov 1;175(4):1141-1155.e16. doi: 
            10.1016/j.cell.2018.09.022. Epub 2018 Oct 18.
  PUBMED    30343902
REFERENCE   2  (bases 1 to 8835)
  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"; doi: 
            "10.1016/j.cell.2018.09.022"; journalName: "Cell"; date: "2018-11-1-
            1"; volume: "175"; issue: "4"; pages: "1141-1155.e16"
COMMENT     Sequence Label: pLKO_HSV.FLAG.NWS_NGFR
FEATURES             Location/Qualifiers
     source          1..8835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..298
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     50..70
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     221..240
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        317..392
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    440..557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        697..1875
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          928..1866
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1823..1843
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1924..1933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2026..2058
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2077..2100
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2122..2145
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    2908..3496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2961..2981)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3379..3390)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3571..3804
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3913..3932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3967..3986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4037..4172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4192..4211)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4193..4211)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4221..4238)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4221..4237)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4230..4252)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4379..4834
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4466..4485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4676..4697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4860..4964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4965..5033,5034..5825)
                     /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(5183..5202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5996..6584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6485..6504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6738..6755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6872..6893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6908..6913,6914..6931,6932..6938)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6946..6962
                     /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     6951..6973
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6970..6986
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6970..6986
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7005..7025
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7007..7025
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7053..7279
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7280..7460
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7507..7632
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8125..8358
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8543..8587
                     /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"
ORIGIN
        1 tcgacggtat cgatcacgag actagtatcg agcggccgcc cccttcaccg agggcctatt
       61 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      121 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      181 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      241 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
      301 gtcttcgaga agacctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      361 atcaacttga aaaagtggca ccgagtcggt gcttttttta attctcgacc tcgagacaaa
      421 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
      481 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
      541 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggccgcggc
      601 tcgaggcgat aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc
      661 taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
      721 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      781 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      841 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
      901 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      961 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
     1021 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
     1081 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
     1141 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
     1201 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
     1261 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
     1321 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     1381 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     1441 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     1501 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     1561 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1621 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     1681 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1741 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1801 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1861 tttcaggtgt cgtgaggaat tctgcagtcg atcgacggta ccgcgggccc gggatccacg
     1921 gtcgccacca tgggggcagg tgccaccggc cgcgccatgg acgggccgcg cctgctgctg
     1981 ttgctgcttc tgggggtgtc ccttggaggt gccaaggagg catgccagcc cgagctggcc
     2041 cccgaggacc ccgaggacgg aggaggcggg ggaggggact acaaagacga tgacgacaag
     2101 ggaggaggcg ggggagggaa ctggtcccac ccccaattcg agaaacccac aggcctgtac
     2161 acacacagcg gtgagtgctg caaagcctgc aacctgggcg agggtgtggc ccagccttgt
     2221 ggagccaacc agaccgtgtg tgagccctgc ctggacagcg tgacgttctc cgacgtggtg
     2281 agcgcgaccg agccgtgcaa gccgtgcacc gagtgcgtgg ggctccagag catgtcggcg
     2341 ccgtgcgtgg aggccgacga cgccgtgtgc cgctgcgcct acggctacta ccaggatgag
     2401 acgactgggc gctgcgaggc gtgccgcgtg tgcgaggcgg gctcgggcct cgtgttctcc
     2461 tgccaggaca agcagaacac cgtgtgcgag gagtgccccg acggcacgta ttccgacgag
     2521 gccaaccacg tggacccgtg cctgccctgc accgtgtgcg aggacaccga gcgccagctc
     2581 cgcgagtgca cacgctgggc cgacgccgag tgcgaggaga tccctggccg ttggattaca
     2641 cggtccacac ccccagaggg ctcggacagc acagccccca gcacccagga gcctgaggca
     2701 cctccagaac aagacctcat agccagcacg gtggcaggtg tggtgaccac agtgatgggc
     2761 agctcccagc ccgtggtgac ccgaggcacc accgacaacc tcatccctgt ctattgctcc
     2821 atcctggctg ctgtggttgt gggccttgtg gcctacatag ccttcaagag gtggaacagg
     2881 ggtatcctct agagcggccg cgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     2941 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3001 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3061 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3121 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3181 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3241 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3301 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3361 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3421 ccggctctgc ggcctcttcc gcatcttcgc cttcgccctc agacgagtcg gatctccctt
     3481 tgggccgcct ccccgcctgg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     3541 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat
     3601 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3781 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     3841 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     3901 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3961 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     4021 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4081 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4141 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     4201 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4261 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4321 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4381 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4441 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4501 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4561 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4621 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4681 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4741 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4801 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4921 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4981 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5041 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5101 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5161 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5221 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5281 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5341 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5401 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5461 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5521 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5701 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6661 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6721 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6781 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6841 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6901 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6961 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     7021 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     7081 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     7141 atggcgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     7201 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     7261 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     7321 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7381 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7441 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     7501 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     7561 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     7621 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     7681 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     7741 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     7801 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     7861 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     7921 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     7981 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     8041 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     8101 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     8161 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     8221 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     8281 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     8341 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     8401 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     8461 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     8521 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8581 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8641 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8701 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     8761 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     8821 attagtgaac ggatc
//
