LOCUS       sequence-327836-        8844 bp    DNA     circular SYN 14-APR-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 8844)
  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 8844)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_HA.V5.FLAG_NGFR
FEATURES             Location/Qualifiers
     source          1..8844
                     /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..2052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2071..2112
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2131..2154
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2917..3505
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2970..2990)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3388..3399)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3580..3813
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3885..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3922..3941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3976..3995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4046..4181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4201..4220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4202..4220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4230..4247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4230..4246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4239..4261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4388..4843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4475..4494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4685..4706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4869..4973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4974..5042,5043..5834)
                     /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(5192..5211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6005..6593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6494..6513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6747..6764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6881..6902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6917..6922,6923..6940,6941..6947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6955..6971
                     /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     6960..6982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6979..6995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6979..6995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7014..7034
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7016..7034
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7062..7288
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7289..7469
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7516..7641
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8134..8367
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8552..8596
                     /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 catgctaccc atacgatgtg
     2041 ccagattacg ctggaggagg cgggggaggg ggcaaaccaa ttccaaaccc actgctggga
     2101 ctggattcca caggaggagg cgggggaggg gactacaaag acgatgacga caagcccaca
     2161 ggcctgtaca cacacagcgg tgagtgctgc aaagcctgca acctgggcga gggtgtggcc
     2221 cagccttgtg gagccaacca gaccgtgtgt gagccctgcc tggacagcgt gacgttctcc
     2281 gacgtggtga gcgcgaccga gccgtgcaag ccgtgcaccg agtgcgtggg gctccagagc
     2341 atgtcggcgc cgtgcgtgga ggccgacgac gccgtgtgcc gctgcgccta cggctactac
     2401 caggatgaga cgactgggcg ctgcgaggcg tgccgcgtgt gcgaggcggg ctcgggcctc
     2461 gtgttctcct gccaggacaa gcagaacacc gtgtgcgagg agtgccccga cggcacgtat
     2521 tccgacgagg ccaaccacgt ggacccgtgc ctgccctgca ccgtgtgcga ggacaccgag
     2581 cgccagctcc gcgagtgcac acgctgggcc gacgccgagt gcgaggagat ccctggccgt
     2641 tggattacac ggtccacacc cccagagggc tcggacagca cagcccccag cacccaggag
     2701 cctgaggcac ctccagaaca agacctcata gccagcacgg tggcaggtgt ggtgaccaca
     2761 gtgatgggca gctcccagcc cgtggtgacc cgaggcacca ccgacaacct catccctgtc
     2821 tattgctcca tcctggctgc tgtggttgtg ggccttgtgg cctacatagc cttcaagagg
     2881 tggaacaggg gtatcctcta gagcggccgc gtcgacaatc aacctctgga ttacaaaatt
     2941 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3001 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3061 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3121 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3181 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3241 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3301 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3361 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3421 ggcctgctgc cggctctgcg gcctcttccg catcttcgcc ttcgccctca gacgagtcgg
     3481 atctcccttt gggccgcctc cccgcctggg tacctttaag accaatgact tacaaggcag
     3541 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     3601 aacgaagata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     3661 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     3721 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3781 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     3841 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     4021 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     4201 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4261 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4321 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4381 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4441 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4501 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4561 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4621 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4681 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4741 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4801 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6901 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6961 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     7021 accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact
     7081 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
     7141 gcaccgtgca tggcgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
     7201 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
     7261 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
     7321 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7381 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7441 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     7501 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     7561 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     7621 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     7681 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     7741 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     7801 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     7861 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     7921 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     7981 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     8041 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     8101 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     8161 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     8221 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     8281 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     8341 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     8401 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     8461 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     8521 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     8581 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     8641 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     8701 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     8761 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
     8821 atccattcga ttagtgaacg gatc
//
