LOCUS       sequence-335195-        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_S.V5.NWS_NGFR
FEATURES             Location/Qualifiers
     source          1..8865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     813..833
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      914..923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1016..1060
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1079..1120
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1142..1165
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1928..2516
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1981..2001)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2399..2410)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2591..2824
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2896..3017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2933..2952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2987..3006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3057..3192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3212..3231)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3213..3231)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3241..3258)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3241..3257)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3250..3272)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3399..3854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3486..3505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3696..3717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3880..3984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3985..4053,4054..4845)
                     /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(4203..4222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5016..5604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5505..5524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5758..5775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5892..5913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5928..5933,5934..5951,5952..5958)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5966..5982
                     /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     5971..5993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5990..6006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5990..6006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6025..6045
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6027..6045
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6073..6299
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6300..6480
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6527..6652
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7145..7378
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7563..7607
                     /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        7905..8153
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7905..7925
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8076..8095
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8172..8247
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    8295..8412
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8552..8865,1..865)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(8783..8865,1..856)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
       61 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
      121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
      181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
      241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
      301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
      361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
      421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
      481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
      541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
      601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
      661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
      721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
      781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
      841 ttttcttcca tttcaggtgt cgtgaggaat tctgcagtcg atcgacggta ccgcgggccc
      901 gggatccacg gtcgccacca tgggggcagg tgccaccggc cgcgccatgg acgggccgcg
      961 cctgctgctg ttgctgcttc tgggggtgtc ccttggaggt gccaaggagg catgcaagga
     1021 gaccgccgcc gccaagttcg agaggcagca catggacagc ggaggaggcg ggggaggggg
     1081 caaaccaatt ccaaacccac tgctgggact ggattccaca ggaggaggcg ggggagggaa
     1141 ctggtcccac ccccaattcg agaaacccac aggcctgtac acacacagcg gtgagtgctg
     1201 caaagcctgc aacctgggcg agggtgtggc ccagccttgt ggagccaacc agaccgtgtg
     1261 tgagccctgc ctggacagcg tgacgttctc cgacgtggtg agcgcgaccg agccgtgcaa
     1321 gccgtgcacc gagtgcgtgg ggctccagag catgtcggcg ccgtgcgtgg aggccgacga
     1381 cgccgtgtgc cgctgcgcct acggctacta ccaggatgag acgactgggc gctgcgaggc
     1441 gtgccgcgtg tgcgaggcgg gctcgggcct cgtgttctcc tgccaggaca agcagaacac
     1501 cgtgtgcgag gagtgccccg acggcacgta ttccgacgag gccaaccacg tggacccgtg
     1561 cctgccctgc accgtgtgcg aggacaccga gcgccagctc cgcgagtgca cacgctgggc
     1621 cgacgccgag tgcgaggaga tccctggccg ttggattaca cggtccacac ccccagaggg
     1681 ctcggacagc acagccccca gcacccagga gcctgaggca cctccagaac aagacctcat
     1741 agccagcacg gtggcaggtg tggtgaccac agtgatgggc agctcccagc ccgtggtgac
     1801 ccgaggcacc accgacaacc tcatccctgt ctattgctcc atcctggctg ctgtggttgt
     1861 gggccttgtg gcctacatag ccttcaagag gtggaacagg ggtatcctct agagcggccg
     1921 cgtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1981 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2041 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2101 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2161 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2221 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2281 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2341 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2401 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2461 gcatcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     2521 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2581 aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt
     2641 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     2701 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2761 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2821 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     2881 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2941 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3001 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     3061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3121 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3181 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     3241 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3301 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3361 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     3421 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3481 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3541 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3601 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3661 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3721 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3781 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3841 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3901 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3961 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4021 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4081 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4141 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4201 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4261 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4321 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4381 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4441 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4501 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4561 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4621 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4681 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4741 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4801 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4861 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4921 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5161 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5701 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5761 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5821 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5881 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5941 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     6001 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     6061 ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga
     6121 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atggcgattg gtggaagtaa
     6181 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa
     6241 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg
     6301 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6361 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6421 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6481 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     6541 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     6601 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     6661 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     6721 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     6781 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     6841 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     6901 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     6961 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     7021 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     7081 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     7141 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     7201 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     7261 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     7321 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     7381 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     7441 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     7501 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     7561 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     7621 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     7681 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     7741 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     7801 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     7861 ggtatcgatc acgagactag tatcgagcgg ccgccccctt caccgagggc ctatttccca
     7921 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     7981 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     8041 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     8101 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggtctt
     8161 cgagaagacc tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     8221 cttgaaaaag tggcaccgag tcggtgcttt ttttaattct cgacctcgag acaaatggca
     8281 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     8341 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     8401 attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag
     8461 gcgataagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
     8521 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     8581 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     8641 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     8701 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     8761 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
     8821 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggct
//
