LOCUS       sequence-329561-        8838 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 8838)
  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 8838)
  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.NWS.HA_NGFR
FEATURES             Location/Qualifiers
     source          1..8838
                     /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             2080..2103
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2122..2148
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2911..3499
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2964..2984)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3382..3393)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3574..3807
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3879..4000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3916..3935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3970..3989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4040..4175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4195..4214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4196..4214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4224..4241)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4224..4240)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4233..4255)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4382..4837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4469..4488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4679..4700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4863..4967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4968..5036,5037..5828)
                     /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(5186..5205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5999..6587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6488..6507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6741..6758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6875..6896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6911..6916,6917..6934,6935..6941)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6949..6965
                     /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     6954..6976
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6973..6989
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6973..6989
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7008..7028
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7010..7028
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7056..7282
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7283..7463
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7510..7635
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8128..8361
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8546..8590
                     /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 ggagggaact ggtcccaccc ccaattcgag
     2101 aaaggaggag gcgggggagg gtacccatac gatgtgccag attacgctcc cacaggcctg
     2161 tacacacaca gcggtgagtg ctgcaaagcc tgcaacctgg gcgagggtgt ggcccagcct
     2221 tgtggagcca accagaccgt gtgtgagccc tgcctggaca gcgtgacgtt ctccgacgtg
     2281 gtgagcgcga ccgagccgtg caagccgtgc accgagtgcg tggggctcca gagcatgtcg
     2341 gcgccgtgcg tggaggccga cgacgccgtg tgccgctgcg cctacggcta ctaccaggat
     2401 gagacgactg ggcgctgcga ggcgtgccgc gtgtgcgagg cgggctcggg cctcgtgttc
     2461 tcctgccagg acaagcagaa caccgtgtgc gaggagtgcc ccgacggcac gtattccgac
     2521 gaggccaacc acgtggaccc gtgcctgccc tgcaccgtgt gcgaggacac cgagcgccag
     2581 ctccgcgagt gcacacgctg ggccgacgcc gagtgcgagg agatccctgg ccgttggatt
     2641 acacggtcca cacccccaga gggctcggac agcacagccc ccagcaccca ggagcctgag
     2701 gcacctccag aacaagacct catagccagc acggtggcag gtgtggtgac cacagtgatg
     2761 ggcagctccc agcccgtggt gacccgaggc accaccgaca acctcatccc tgtctattgc
     2821 tccatcctgg ctgctgtggt tgtgggcctt gtggcctaca tagccttcaa gaggtggaac
     2881 aggggtatcc tctagagcgg ccgcgtcgac aatcaacctc tggattacaa aatttgtgaa
     2941 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3001 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3061 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3121 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3181 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3241 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3301 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3361 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3421 ctgccggctc tgcggcctct tccgcatctt cgccttcgcc ctcagacgag tcggatctcc
     3481 ctttgggccg cctccccgcc tgggtacctt taagaccaat gacttacaag gcagctgtag
     3541 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     3601 gataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     3661 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     3721 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     3781 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     3841 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     3901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     4021 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4081 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4141 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     4201 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4261 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4321 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4381 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4441 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4501 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4561 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4621 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4681 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4741 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4801 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     4861 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4921 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4981 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5041 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5101 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5161 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5221 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5281 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5341 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5401 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5461 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5521 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5581 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5641 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5701 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5761 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5821 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5881 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5941 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6781 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6841 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6901 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6961 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     7021 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
     7081 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
     7141 tgcatggcga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
     7201 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
     7261 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
     7321 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7381 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7441 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     7501 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     7561 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     7621 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     7681 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     7741 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     7801 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     7861 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     7921 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     7981 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     8041 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     8101 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     8161 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     8221 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     8281 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     8341 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     8401 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     8461 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     8521 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     8581 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     8641 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     8701 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     8761 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     8821 tcgattagtg aacggatc
//
