LOCUS       sequence-330661-        8853 bp    DNA     circular SYN 15-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 8853)
  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 8853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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.FLAG.VSVg_NGFR
FEATURES             Location/Qualifiers
     source          1..8853
                     /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..2070
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2089..2112
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2131..2163
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    2926..3514
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2979..2999)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3397..3408)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3589..3822
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3894..4015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3931..3950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3985..4004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4055..4190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4210..4229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4211..4229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4239..4256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4239..4255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4248..4270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4397..4852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4484..4503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4694..4715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4983..5051,5052..5843)
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6756..6773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6890..6911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6926..6931,6932..6949,6950..6956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6964..6980
                     /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     6969..6991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6988..7004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6988..7004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7023..7043
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7025..7043
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7071..7297
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7298..7478
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7525..7650
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8143..8376
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8561..8605
                     /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 catgcaagga gaccgccgcc
     2041 gccaagttcg agaggcagca catggacagc ggaggaggcg ggggagggga ctacaaagac
     2101 gatgacgaca agggaggagg cgggggaggg tatacagata tcgaaatgaa tagactcgga
     2161 aagcccacag gcctgtacac acacagcggt gagtgctgca aagcctgcaa cctgggcgag
     2221 ggtgtggccc agccttgtgg agccaaccag accgtgtgtg agccctgcct ggacagcgtg
     2281 acgttctccg acgtggtgag cgcgaccgag ccgtgcaagc cgtgcaccga gtgcgtgggg
     2341 ctccagagca tgtcggcgcc gtgcgtggag gccgacgacg ccgtgtgccg ctgcgcctac
     2401 ggctactacc aggatgagac gactgggcgc tgcgaggcgt gccgcgtgtg cgaggcgggc
     2461 tcgggcctcg tgttctcctg ccaggacaag cagaacaccg tgtgcgagga gtgccccgac
     2521 ggcacgtatt ccgacgaggc caaccacgtg gacccgtgcc tgccctgcac cgtgtgcgag
     2581 gacaccgagc gccagctccg cgagtgcaca cgctgggccg acgccgagtg cgaggagatc
     2641 cctggccgtt ggattacacg gtccacaccc ccagagggct cggacagcac agcccccagc
     2701 acccaggagc ctgaggcacc tccagaacaa gacctcatag ccagcacggt ggcaggtgtg
     2761 gtgaccacag tgatgggcag ctcccagccc gtggtgaccc gaggcaccac cgacaacctc
     2821 atccctgtct attgctccat cctggctgct gtggttgtgg gccttgtggc ctacatagcc
     2881 ttcaagaggt ggaacagggg tatcctctag agcggccgcg tcgacaatca acctctggat
     2941 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3001 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3061 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3121 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3181 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3241 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3301 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     3361 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3421 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc atcttcgcct tcgccctcag
     3481 acgagtcgga tctccctttg ggccgcctcc ccgcctgggt acctttaaga ccaatgactt
     3541 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     3601 ttcactccca acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     3661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     3721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     3781 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     3841 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     3901 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3961 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4021 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4081 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4141 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     4201 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     4261 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4321 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4381 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4441 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4501 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4561 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4621 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4681 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4741 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4801 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6781 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6841 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6901 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6961 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7021 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     7081 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     7141 agagaaaaag caccgtgcat ggcgattggt ggaagtaagg tggtacgatc gtgccttatt
     7201 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     7261 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7321 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7381 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7441 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7501 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7561 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7621 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7681 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7741 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7801 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7861 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7921 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7981 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     8041 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     8101 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     8161 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     8221 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8281 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8341 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8401 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8461 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8521 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8581 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8641 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8701 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8761 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     8821 cagagacaga tccattcgat tagtgaacgg atc
//
