LOCUS       sequence-305412-        8855 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 8855)
  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 8855)
  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.Ollas.HA_NGFR
FEATURES             Location/Qualifiers
     source          1..8855
                     /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..1874
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          928..1865
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1822..1842
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1923..1932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2025..2057
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2076..2117
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2139..2165
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2928..3516
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2981..3001)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3399..3410)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3591..3824
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3896..4017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3933..3952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3987..4006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4057..4192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4212..4231)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4213..4231)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4241..4258)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4241..4257)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4250..4272)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4399..4854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4486..4505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4696..4717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4880..4984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4985..5053,5054..5845)
                     /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(5203..5222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6016..6604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6505..6524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6758..6775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6892..6913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6928..6933,6934..6951,6952..6958)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6966..6982
                     /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     6971..6993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6990..7006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6990..7006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7025..7045
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7027..7045
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7073..7299
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7300..7480
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7527..7652
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8145..8378
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8563..8607
                     /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 ccactggctg cagtacgtga
     1021 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     1081 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     1141 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     1201 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     1261 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     1321 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     1381 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     1441 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     1501 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     1561 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1621 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1681 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1741 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1801 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1861 ttcaggtgtc gtgaggaatt ctgcagtcga tcgacggtac cgcgggcccg ggatccacgg
     1921 tcgccaccat gggggcaggt gccaccggcc gcgccatgga cgggccgcgc ctgctgctgt
     1981 tgctgcttct gggggtgtcc cttggaggtg ccaaggaggc atgccagccc gagctggccc
     2041 ccgaggaccc cgaggacgga ggaggcgggg gagggagcgg cttcgccaac gagctgggcc
     2101 ccaggctgat gggcaagtgc ggaggaggcg ggggagggta cccatacgat gtgccagatt
     2161 acgctcccac aggcctgtac acacacagcg gtgagtgctg caaagcctgc aacctgggcg
     2221 agggtgtggc ccagccttgt ggagccaacc agaccgtgtg tgagccctgc ctggacagcg
     2281 tgacgttctc cgacgtggtg agcgcgaccg agccgtgcaa gccgtgcacc gagtgcgtgg
     2341 ggctccagag catgtcggcg ccgtgcgtgg aggccgacga cgccgtgtgc cgctgcgcct
     2401 acggctacta ccaggatgag acgactgggc gctgcgaggc gtgccgcgtg tgcgaggcgg
     2461 gctcgggcct cgtgttctcc tgccaggaca agcagaacac cgtgtgcgag gagtgccccg
     2521 acggcacgta ttccgacgag gccaaccacg tggacccgtg cctgccctgc accgtgtgcg
     2581 aggacaccga gcgccagctc cgcgagtgca cacgctgggc cgacgccgag tgcgaggaga
     2641 tccctggccg ttggattaca cggtccacac ccccagaggg ctcggacagc acagccccca
     2701 gcacccagga gcctgaggca cctccagaac aagacctcat agccagcacg gtggcaggtg
     2761 tggtgaccac agtgatgggc agctcccagc ccgtggtgac ccgaggcacc accgacaacc
     2821 tcatccctgt ctattgctcc atcctggctg ctgtggttgt gggccttgtg gcctacatag
     2881 ccttcaagag gtggaacagg ggtatcctct agagcggccg cgtcgacaat caacctctgg
     2941 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3001 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3061 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3121 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3181 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3241 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3301 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     3361 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3421 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcatcttcgc cttcgccctc
     3481 agacgagtcg gatctccctt tgggccgcct ccccgcctgg gtacctttaa gaccaatgac
     3541 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     3601 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga
     3661 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     3721 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     3781 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     3841 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     3901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4021 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4081 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4141 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     4201 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     4261 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4321 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4381 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4861 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5221 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5521 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6781 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6841 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6901 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6961 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     7021 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
     7081 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
     7141 ggagagaaaa agcaccgtgc atggcgattg gtggaagtaa ggtggtacga tcgtgcctta
     7201 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
     7261 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
     7321 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7381 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7441 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     7501 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     7561 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     7621 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     7681 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     7741 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     7801 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     7861 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     7921 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     7981 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     8041 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     8101 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     8161 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     8221 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     8281 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     8341 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     8401 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     8461 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     8521 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     8581 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     8641 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     8701 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     8761 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     8821 gacagagaca gatccattcg attagtgaac ggatc
//
