LOCUS       sequence-329410-        8841 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 8841)
  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 8841)
  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.FLAG.VSVg_NGFR
FEATURES             Location/Qualifiers
     source          1..8841
                     /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..2100
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2119..2151
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    2914..3502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2967..2987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3385..3396)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3577..3810
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3882..4003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3919..3938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3973..3992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4198..4217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4199..4217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4227..4244)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4227..4243)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4258)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4385..4840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4472..4491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4682..4703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4866..4970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4971..5039,5040..5831)
                     /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(5189..5208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6002..6590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6491..6510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6744..6761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6878..6899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6914..6919,6920..6937,6938..6944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6952..6968
                     /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     6957..6979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6976..6992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6976..6992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7011..7031
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7013..7031
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7059..7285
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7286..7466
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7513..7638
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8131..8364
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8549..8593
                     /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 ggaggggact acaaagacga tgacgacaag
     2101 ggaggaggcg ggggagggta tacagatatc gaaatgaata gactcggaaa gcccacaggc
     2161 ctgtacacac acagcggtga gtgctgcaaa gcctgcaacc tgggcgaggg tgtggcccag
     2221 ccttgtggag ccaaccagac cgtgtgtgag ccctgcctgg acagcgtgac gttctccgac
     2281 gtggtgagcg cgaccgagcc gtgcaagccg tgcaccgagt gcgtggggct ccagagcatg
     2341 tcggcgccgt gcgtggaggc cgacgacgcc gtgtgccgct gcgcctacgg ctactaccag
     2401 gatgagacga ctgggcgctg cgaggcgtgc cgcgtgtgcg aggcgggctc gggcctcgtg
     2461 ttctcctgcc aggacaagca gaacaccgtg tgcgaggagt gccccgacgg cacgtattcc
     2521 gacgaggcca accacgtgga cccgtgcctg ccctgcaccg tgtgcgagga caccgagcgc
     2581 cagctccgcg agtgcacacg ctgggccgac gccgagtgcg aggagatccc tggccgttgg
     2641 attacacggt ccacaccccc agagggctcg gacagcacag cccccagcac ccaggagcct
     2701 gaggcacctc cagaacaaga cctcatagcc agcacggtgg caggtgtggt gaccacagtg
     2761 atgggcagct cccagcccgt ggtgacccga ggcaccaccg acaacctcat ccctgtctat
     2821 tgctccatcc tggctgctgt ggttgtgggc cttgtggcct acatagcctt caagaggtgg
     2881 aacaggggta tcctctagag cggccgcgtc gacaatcaac ctctggatta caaaatttgt
     2941 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3001 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3061 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3121 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3181 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3241 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3301 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3361 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3421 ctgctgccgg ctctgcggcc tcttccgcat cttcgccttc gccctcagac gagtcggatc
     3481 tccctttggg ccgcctcccc gcctgggtac ctttaagacc aatgacttac aaggcagctg
     3541 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     3601 gaagataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     3661 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     3721 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     3781 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     3841 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     3901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     4021 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     4201 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     4261 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4321 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4381 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     4861 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4921 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4981 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5041 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5101 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5161 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5221 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5281 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5341 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5401 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5461 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5521 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5581 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5641 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5701 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5761 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5821 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5881 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5941 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     6841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     6901 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     6961 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     7021 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
     7081 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     7141 ccgtgcatgg cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     7201 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     7261 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     7321 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     7381 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     7441 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     7501 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     7561 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     7621 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     7681 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     7741 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     7801 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     7861 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     7921 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     7981 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     8041 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     8101 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     8161 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     8221 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     8281 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     8341 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     8401 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     8461 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     8521 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     8581 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     8641 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     8701 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     8761 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     8821 cattcgatta gtgaacggat c
//
