LOCUS       sequence-335191-        8862 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 8862)
  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 8862)
  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_Ollas.V5.FLAG_NGFR
FEATURES             Location/Qualifiers
     source          1..8862
                     /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..2067
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2089..2130
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2149..2172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2935..3523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2988..3008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3406..3417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3598..3831
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3903..4024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3940..3959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3994..4013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4064..4199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4219..4238)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4220..4238)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4248..4265)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4248..4264)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4257..4279)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4406..4861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4703..4724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4887..4991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4992..5060,5061..5852)
                     /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(5210..5229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6023..6611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6512..6531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6765..6782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6899..6920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6935..6940,6941..6958,6959..6965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6973..6989
                     /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     6978..7000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6997..7013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6997..7013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7032..7052
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7034..7052
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7080..7306
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7307..7487
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7534..7659
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8152..8385
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8570..8614
                     /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 catgcagcgg cttcgccaac
     2041 gagctgggcc ccaggctgat gggcaagtgc ggaggaggcg ggggaggggg caaaccaatt
     2101 ccaaacccac tgctgggact ggattccaca ggaggaggcg ggggagggga ctacaaagac
     2161 gatgacgaca agcccacagg cctgtacaca cacagcggtg agtgctgcaa agcctgcaac
     2221 ctgggcgagg gtgtggccca gccttgtgga gccaaccaga ccgtgtgtga gccctgcctg
     2281 gacagcgtga cgttctccga cgtggtgagc gcgaccgagc cgtgcaagcc gtgcaccgag
     2341 tgcgtggggc tccagagcat gtcggcgccg tgcgtggagg ccgacgacgc cgtgtgccgc
     2401 tgcgcctacg gctactacca ggatgagacg actgggcgct gcgaggcgtg ccgcgtgtgc
     2461 gaggcgggct cgggcctcgt gttctcctgc caggacaagc agaacaccgt gtgcgaggag
     2521 tgccccgacg gcacgtattc cgacgaggcc aaccacgtgg acccgtgcct gccctgcacc
     2581 gtgtgcgagg acaccgagcg ccagctccgc gagtgcacac gctgggccga cgccgagtgc
     2641 gaggagatcc ctggccgttg gattacacgg tccacacccc cagagggctc ggacagcaca
     2701 gcccccagca cccaggagcc tgaggcacct ccagaacaag acctcatagc cagcacggtg
     2761 gcaggtgtgg tgaccacagt gatgggcagc tcccagcccg tggtgacccg aggcaccacc
     2821 gacaacctca tccctgtcta ttgctccatc ctggctgctg tggttgtggg ccttgtggcc
     2881 tacatagcct tcaagaggtg gaacaggggt atcctctaga gcggccgcgt cgacaatcaa
     2941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3301 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     3361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgca tcttcgcctt
     3481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgggta cctttaagac
     3541 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     3601 aagggctaat tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct
     3661 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     3721 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     3781 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     3841 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     3901 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3961 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4021 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     4081 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4141 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4201 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     4261 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4321 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4381 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4621 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4741 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4861 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4921 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5221 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5341 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5521 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5581 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5821 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5881 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6001 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6901 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7021 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     7081 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     7141 cttacaagga gagaaaaagc accgtgcatg gcgattggtg gaagtaaggt ggtacgatcg
     7201 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     7261 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     7321 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     7381 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     7441 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     7501 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     7561 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     7621 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     7681 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     7741 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     7801 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     7861 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     7921 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     7981 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     8041 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     8101 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     8161 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     8221 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     8281 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     8341 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     8401 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     8461 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     8521 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     8581 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     8641 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     8701 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     8761 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     8821 agagagagac agagacagat ccattcgatt agtgaacgga tc
//
