LOCUS       sequence-330447-        8856 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 8856)
  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 8856)
  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_NWS.Ollas.VSVg_NGFR
FEATURES             Location/Qualifiers
     source          1..8856
                     /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             1019..1042
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1061..1102
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             1124..1156
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    1919..2507
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1972..1992)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2390..2401)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2582..2815
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2887..3008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2924..2943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2978..2997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3048..3183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3203..3222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3204..3222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3232..3249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3232..3248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3241..3263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3390..3845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3477..3496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3687..3708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3871..3975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3976..4044,4045..4836)
                     /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(4194..4213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5007..5595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5496..5515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5749..5766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5883..5904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5919..5924,5925..5942,5943..5949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5957..5973
                     /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     5962..5984
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5981..5997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5981..5997
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6016..6036
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6018..6036
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6064..6290
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6291..6471
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6518..6643
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7136..7369
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7554..7598
                     /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        7896..8144
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7896..7916
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8067..8086
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8163..8238
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    8286..8403
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8543..8856,1..865)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(8774..8856,1..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 catgcaactg
     1021 gtcccacccc caattcgaga aaggaggagg cgggggaggg agcggcttcg ccaacgagct
     1081 gggccccagg ctgatgggca agtgcggagg aggcggggga gggtatacag atatcgaaat
     1141 gaatagactc ggaaagccca caggcctgta cacacacagc ggtgagtgct gcaaagcctg
     1201 caacctgggc gagggtgtgg cccagccttg tggagccaac cagaccgtgt gtgagccctg
     1261 cctggacagc gtgacgttct ccgacgtggt gagcgcgacc gagccgtgca agccgtgcac
     1321 cgagtgcgtg gggctccaga gcatgtcggc gccgtgcgtg gaggccgacg acgccgtgtg
     1381 ccgctgcgcc tacggctact accaggatga gacgactggg cgctgcgagg cgtgccgcgt
     1441 gtgcgaggcg ggctcgggcc tcgtgttctc ctgccaggac aagcagaaca ccgtgtgcga
     1501 ggagtgcccc gacggcacgt attccgacga ggccaaccac gtggacccgt gcctgccctg
     1561 caccgtgtgc gaggacaccg agcgccagct ccgcgagtgc acacgctggg ccgacgccga
     1621 gtgcgaggag atccctggcc gttggattac acggtccaca cccccagagg gctcggacag
     1681 cacagccccc agcacccagg agcctgaggc acctccagaa caagacctca tagccagcac
     1741 ggtggcaggt gtggtgacca cagtgatggg cagctcccag cccgtggtga cccgaggcac
     1801 caccgacaac ctcatccctg tctattgctc catcctggct gctgtggttg tgggccttgt
     1861 ggcctacata gccttcaaga ggtggaacag gggtatcctc tagagcggcc gcgtcgacaa
     1921 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1981 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2041 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2101 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2161 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2221 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2281 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2341 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2401 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcatcttcg
     2461 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg ggtaccttta
     2521 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     2581 ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg tactgggtct
     2641 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     2701 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     2761 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     2821 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     2881 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2941 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3001 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     3061 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3181 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     3241 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3301 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3361 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     3421 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3481 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3541 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3601 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3661 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3721 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3781 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3841 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3901 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3961 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4021 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4081 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4141 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4201 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4261 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4321 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4381 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4441 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4501 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4561 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4621 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4681 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4741 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4801 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4861 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4921 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4981 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5041 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5101 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5161 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5221 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5281 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5341 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5401 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5461 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5521 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5581 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5641 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5701 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5761 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5821 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5881 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     5941 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6001 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct
     6061 taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac
     6121 atgccttaca aggagagaaa aagcaccgtg catggcgatt ggtggaagta aggtggtacg
     6181 atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga accactgaat
     6241 tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct
     6301 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6361 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6421 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     6481 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     6541 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     6601 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     6661 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     6721 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     6781 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     6841 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     6901 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     6961 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     7021 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     7081 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     7141 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     7201 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     7261 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     7321 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     7381 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     7441 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     7501 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     7561 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     7621 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     7681 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     7741 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     7801 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat
     7861 cacgagacta gtatcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt
     7921 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
     7981 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     8041 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     8101 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggtct tcgagaagac
     8161 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     8221 gtggcaccga gtcggtgctt tttttaattc tcgacctcga gacaaatggc agtattcatc
     8281 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     8341 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     8401 tttcgggttt attacaggga cagcagagat ccactttggc cgcggctcga ggcgataagc
     8461 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
     8521 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     8581 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     8641 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     8701 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     8761 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     8821 tggcccttgc gtgccttgaa ttacttccac ctggct
//
