LOCUS       sequence-222766-        8902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA Ef1alpha Puro-T2A-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8902)
  AUTHORS   Horlbeck MA, Xu A, Wang M, Bennett NK, Park CY, Bogdanoff D, Adamson
            B, Chow ED, Kampmann M, Peterson TR, Nakamura K, Fischbach MA, 
            Weissman JS, Gilbert LA
  TITLE     Mapping the Genetic Landscape of Human Cells.
  JOURNAL   Cell. 2018 Aug 9;174(4):953-967.e22. doi: 
            10.1016/j.cell.2018.06.010. Epub 2018 Jul 19.
  PUBMED    30033366
REFERENCE   2  (bases 1 to 8902)
  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.2018.06.010"; journalName: "Cell"; date: "2018-08-9-
            9"; volume: "174"; issue: "4"; pages: "953-967.e22"
COMMENT     Sequence Label: pU6-sgRNA Ef1alpha Puro-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..8902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             836..1016
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1063..1188
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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"
     misc_feature    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2815..2848)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2852..2872
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     promoter        3118..3329
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          3330..4268
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4225..4245
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4289..4885
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4289..4308)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4785..4805
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4895..4948
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      4955..4964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4961..4963,4964..4966,4967..5680)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5006..5027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5267..5286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5614..5635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5703..5736)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5792..6380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5845..5865)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6263..6274)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6383..6399)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6384..6400)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6909..7089
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7151..7739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7231..7250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7910..8701,8702..8770))
                     /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     8533..8552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8771..8875)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
      841 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      901 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      961 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     1021 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     1081 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     1141 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     1201 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     1261 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     1321 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     1381 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     1441 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     1501 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgctctaga gatccgacgc gccatctcta
     2641 ggcccgcgcc ggccccctcg cacggacttg tgggagaagc tcggctactc ccctgccccg
     2701 gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga taaaagacag
     2761 ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat ttctataact
     2821 tcgtatagca tacattatac gaagttataa acagcacaaa aggaaactca ccctaactgt
     2881 aaagtaattg tgtgttttga gactataagt atcccttgga gaaccacctt gttggaacga
     2941 ctagttaggc gtgtagttta agagctaagc tggaaacagc atagcaagtt taaataaggc
     3001 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttct cgagtactag
     3061 gatccattag gcggccgcgt ggataaccgt attaccgcca tgcatgtgcc cgtcagtggg
     3121 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     3181 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     3241 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     3301 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     3361 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     3421 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     3481 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     3541 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     3601 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     3661 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     3721 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     3781 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     3841 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     3901 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     3961 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     4021 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgcgc ttttggagta
     4081 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     4141 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     4201 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     4261 catttcaggt gtcgtgagct agcccaccat gaccgagtac aagcccacgg tgcgcctcgc
     4321 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4381 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4441 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4501 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4561 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4621 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4681 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4741 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4801 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4861 gtgcatgacc cgcaagcccg gtgccggatc gggagagggc agaggaagtc tgctaacatg
     4921 cggtgacgtc gaggagaatc ctggcccacc ggtcgccacc atggtgagca agggcgagga
     4981 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     5041 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     5101 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     5161 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     5221 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     5281 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5341 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     5401 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa
     5461 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     5521 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     5581 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5641 cgccgggatc actctcggca tggacgagct gtacaagtaa taagaattcg tcgagggacc
     5701 taataacttc gtatagcata cattatacga agttatacat gtttaagggt tccggttcca
     5761 ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     5821 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5881 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5941 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6001 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6061 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6121 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6181 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6241 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6301 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6361 cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg
     6421 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     6481 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     6541 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6601 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     6661 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
     6721 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
     6781 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
     6841 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
     6901 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6961 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     7021 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     7081 tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8221 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8521 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8881 ttccccgaaa agtgccacct ga
//
