LentiGuide Blast

LOCUS       LentiGuide Blast        9981 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and blasticidin resistance from EF-1a 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9981)
  AUTHORS   Luo R, Yan J, Oh JW, Xi W, Shigaki D, Wong W, Cho HS, Murphy D, 
            Cutler R, Rosen BP, Pulecio J, Yang D, Glenn RA, Chen T, Li QV, 
            Vierbuchen T, Sidoli S, Apostolou E, Huangfu D, Beer MA
  TITLE     Dynamic network-guided CRISPRi screen identifies 
            CTCF-loop-constrained nonlinear enhancer gene regulatory activity 
            during cell state transitions.
  JOURNAL   Nat Genet. 2023 Aug;55(8):1336-1346. doi: 
            10.1038/s41588-023-01450-7. Epub 2023 Jul 24.
  PUBMED    37488417
REFERENCE   2  (bases 1 to 9981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41588-023-01450-7"; journalName: "Nat Genet"; date: 
            "2023-08"; volume: "55"; issue: "8"; pages: "1336-1346"
COMMENT     Sequence Label: LentiGuide Blast
FEATURES             Location/Qualifiers
     source          1..9981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        162..410
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     162..182
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     333..352
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2292..2367
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2423..2540
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2679..3857
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2910..3848
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3805..3825
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3864..3873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3870..4268
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    4283..4871
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4336..4356)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4754..4765)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4943..5176
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5254..5375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5291..5310)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5345..5364
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5415..5550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5570..5589)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5571..5589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5599..5616)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5599..5615)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5608..5630)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5757..6212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5844..5863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6054..6075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6238..6342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6343..6411,6412..7203)
                     /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(6561..6580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7374..7962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7863..7882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8116..8133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8250..8271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8286..8291,8292..8309,8310..8316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8324..8340
                     /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     8329..8351
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8348..8364
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8348..8364
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8383..8403
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8385..8403
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8431..8657
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8658..8838
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8885..9010
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9503..9736
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9921..9965
                     /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 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
       61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      121 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
      181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
      241 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
      301 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
      361 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt
      421 gtaaatgagc acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc
      481 aaccaaaatc ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt
      541 tgcaaacttt taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg
      601 ttttcttcac ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct
      661 tcaatcattt tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca
      721 ttgtctttaa caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa
      781 aatcctgttg atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc
      841 atgatttctt cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca
      901 gctgcttttt ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat
      961 gcaccaatat ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact
     1021 tcgaatgctt ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt
     1081 cctacatttg tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa
     1141 actgttgcaa aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta
     1201 gcatcttgtt ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata
     1261 tattcatttt taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa
     1321 cagattcacc aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa
     1381 cagcaaatca tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat
     1441 acattccatc gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg
     1501 gtgttgctct caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa
     1561 aattcaagtt taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat
     1621 agtctactaa tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag
     1681 aagcttgtat tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt
     1741 cataagcaaa aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc
     1801 atttaaatcc agttaaagtt tttacaatat taactccata tttttcatga gcgattctat
     1861 cacccaaatc acttgttaca aaacttgaat atagagccgg attttttgga atgctattta
     1921 agcgttttag atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta
     1981 atcttacaaa atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat
     2041 tcataataat aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag
     2101 gatcaaattc tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct
     2161 cttcaacctc aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc
     2221 ctgttccatt aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt
     2281 tgtacgtctc tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     2341 cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag atcttgagac
     2401 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2461 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2521 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
     2581 ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca
     2641 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2701 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2761 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2821 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2881 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2941 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     3001 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     3061 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     3121 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     3181 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     3241 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     3301 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     3361 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     3421 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     3481 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     3541 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     3601 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     3661 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3721 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3781 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3841 catttcaggt gtcgtgacgt acggccacca tggccaagcc tttgtctcaa gaagaatcca
     3901 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     3961 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     4021 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     4081 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     4141 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     4201 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     4261 agggctaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4321 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4381 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4441 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4501 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4561 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4621 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4681 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4741 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4801 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4861 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4921 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     4981 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5041 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5101 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5161 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     5221 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5281 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5341 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     5401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5521 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     5581 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5641 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5701 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6181 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6241 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6301 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6361 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6421 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6481 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6541 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6601 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6661 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6721 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6781 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6841 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6901 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6961 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7021 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7081 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7141 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7201 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7261 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7321 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7381 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7441 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7501 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7561 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7621 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7681 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7741 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7801 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7861 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7921 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7981 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8041 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8101 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8161 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8221 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8281 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8341 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     8401 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     8461 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     8521 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     8581 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     8641 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8701 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8761 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8821 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     8881 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     8941 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     9001 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     9061 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     9121 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     9181 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     9241 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     9301 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     9361 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     9421 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     9481 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     9541 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     9601 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     9661 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     9721 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     9781 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     9841 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     9901 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     9961 gtttgtggaa ttggtttaac a
//