LOCUS       sequence-328342-        8865 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 8865)
  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 8865)
  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.S.C_NGFR
FEATURES             Location/Qualifiers
     source          1..8865
                     /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..2121
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2140..2175
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     misc_feature    2938..3526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2991..3011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3409..3420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3601..3834
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3906..4027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3943..3962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3997..4016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4067..4202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4222..4241)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4223..4241)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4251..4268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4251..4267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4260..4282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4409..4864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4496..4515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4706..4727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4890..4994
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4995..5063,5064..5855)
                     /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(5213..5232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6026..6614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6515..6534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6768..6785
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6902..6923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6938..6943,6944..6961,6962..6968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6976..6992
                     /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     6981..7003
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7000..7016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7000..7016
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7035..7055
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7037..7055
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7083..7309
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7310..7490
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7537..7662
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8155..8388
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8573..8617
                     /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 ggagggaagg agaccgccgc cgccaagttc
     2101 gagaggcagc acatggacag cggaggaggc gggggagggg aggaccaggt ggaccccagg
     2161 ctgatcgacg gcaagcccac aggcctgtac acacacagcg gtgagtgctg caaagcctgc
     2221 aacctgggcg agggtgtggc ccagccttgt ggagccaacc agaccgtgtg tgagccctgc
     2281 ctggacagcg tgacgttctc cgacgtggtg agcgcgaccg agccgtgcaa gccgtgcacc
     2341 gagtgcgtgg ggctccagag catgtcggcg ccgtgcgtgg aggccgacga cgccgtgtgc
     2401 cgctgcgcct acggctacta ccaggatgag acgactgggc gctgcgaggc gtgccgcgtg
     2461 tgcgaggcgg gctcgggcct cgtgttctcc tgccaggaca agcagaacac cgtgtgcgag
     2521 gagtgccccg acggcacgta ttccgacgag gccaaccacg tggacccgtg cctgccctgc
     2581 accgtgtgcg aggacaccga gcgccagctc cgcgagtgca cacgctgggc cgacgccgag
     2641 tgcgaggaga tccctggccg ttggattaca cggtccacac ccccagaggg ctcggacagc
     2701 acagccccca gcacccagga gcctgaggca cctccagaac aagacctcat agccagcacg
     2761 gtggcaggtg tggtgaccac agtgatgggc agctcccagc ccgtggtgac ccgaggcacc
     2821 accgacaacc tcatccctgt ctattgctcc atcctggctg ctgtggttgt gggccttgtg
     2881 gcctacatag ccttcaagag gtggaacagg ggtatcctct agagcggccg cgtcgacaat
     2941 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3001 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3061 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3121 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3181 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3241 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3301 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3361 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3421 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcatcttcgc
     3481 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg gtacctttaa
     3541 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     3601 tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc
     3661 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     3721 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     3781 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     3841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     3901 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4021 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     4081 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4141 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4201 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     4261 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4321 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4381 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4621 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4741 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4801 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4861 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4921 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4981 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5041 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5101 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5161 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5221 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5281 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5341 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5401 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5461 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5521 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5581 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5641 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5701 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5761 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5821 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5881 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5941 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6001 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6061 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6541 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6601 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6661 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6721 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6781 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6841 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6901 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6961 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7021 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     7081 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     7141 tgccttacaa ggagagaaaa agcaccgtgc atggcgattg gtggaagtaa ggtggtacga
     7201 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     7261 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     7321 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7381 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7441 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     7501 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     7561 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     7621 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     7681 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     7741 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     7801 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     7861 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     7921 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     7981 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     8041 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     8101 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     8161 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     8221 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     8281 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     8341 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     8401 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     8461 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     8521 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     8581 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     8641 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     8701 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     8761 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     8821 tggagagaga gacagagaca gatccattcg attagtgaac ggatc
//
