LOCUS       sequence-430620-       11391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and hygromycin resistance marker with 2A 
            mTagBFP2 from EF-1a promoter. 3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11391)
  AUTHORS   Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, 
            Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ
  TITLE     Evaluating Synthetic Activation and Repression of 
            Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and 
            Astrocytes.
  JOURNAL   Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 
            10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.
  PUBMED    28757163
REFERENCE   2  (bases 1 to 11391)
  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.stemcr.2017.06.012"; journalName: "Stem Cell Reports"; 
            date: "2017-08-8- 8"; volume: "9"; issue: "2"; pages: "615-628"
COMMENT     Sequence Label: lentiGuide-Hygro-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..11391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(301..549)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(359..378)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(529..549)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(727..771)
                     /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    956..1189
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1682..1807
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1854..2034
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2035..2261)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2289..2309)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2289..2307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2328..2344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2328..2344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2341..2363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2352..2368
                     /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)."
     promoter        complement(join(2376..2382,2383..2400,2401..2406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2421..2442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2559..2576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2730..3318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2810..2829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3489..4280,4281..4349))
                     /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     4112..4131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4350..4454)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4480..4935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4829..4848
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5062..5084
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5076..5093
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5077..5093
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5103..5122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5103..5121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      5142..5277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5317..5438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5328..5347)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5382..5401
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5516..5749
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    5821..6409
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5927..5938
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6336..6356
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6424..7125)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             complement(7126..7182)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(7207..8223)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     regulatory      8229..8238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8245..9423)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8254..9192
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(8277..8297)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    9562..9679
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(9735..9810)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 aaagcagctg gtggagttag aacatttgaa gatgctcaaa aaatgattga agcaggagct
       61 gaccgcttag gaacaagtgg tggagtagct attattaaag gtgaagaaaa caacgcgagt
      121 tactaaaact agcgtttttt tattttgctc atttttatta aaagtttgca aaaaggaaca
      181 taaaaattct aattattgat actaaagtta ttaaaaagaa gattttggtt gattttataa
      241 aggtcataga atataatatt ttagcatgtg tattttgtgt gctcatttac aaccgtctcc
      301 ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac
      361 ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa
      421 ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag tcaaattaat
      481 tctaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa
      541 taggccctct ctgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
      601 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
      661 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
      721 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
      781 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
      841 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
      901 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
      961 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     1021 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     1081 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     1141 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     1201 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     1261 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     1321 tccaggtctg aagatcagcg gccgcttgct gtgcggtggt cttacttttg ttttgctctt
     1381 cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat
     1441 agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg
     1501 gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccaggattaa
     1561 ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct
     1621 ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac
     1681 tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac
     1741 tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc
     1801 gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga
     1861 gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag
     1921 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
     1981 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccgtttat
     2041 gtatcgagct aggcacttaa atacaatatc tctgcaatgc ggcaattcag tggttcgtcc
     2101 aatccatgtc agacccgtct gttgccttcc taataaggca cgatcgtacc accttacttc
     2161 caccaatcgg catgcacggt gctttttctc tccttgtaag gcatgttgct aactcatcgt
     2221 taccatgttg caagactaca agagtattgc ataagactac attaagcttg cagctccagc
     2281 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2341 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2401 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2521 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2581 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2641 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2701 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2761 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2821 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2881 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2941 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3001 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3061 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3121 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3181 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3241 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3361 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4441 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4501 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4561 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4621 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4681 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4741 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4801 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4861 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4921 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4981 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     5041 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     5101 cgtaatacga ctcactatag ggcgaattgg gtacgtccct aggcctccaa aaaagcctcc
     5161 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     5221 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     5281 cggagttagg ggcgggatag ctagagccag acatgataag atacattgat gagtttggac
     5341 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     5401 ctttatttgt aaccattata agctgcaata aacaagttcc tctcactctc tgatattcat
     5461 ttctttgcaa gttataaata ctgaataata agatgacatg aactactata cgtactgcta
     5521 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     5581 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     5641 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     5701 caagcaaaaa gcagatcttg tcttcgttgg gagtgaatta gcccttccag tccccccttt
     5761 tcttttaaaa agtggctaag atctacagct gccttgtaag tcattggtct taaagtcgac
     5821 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     5881 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     5941 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     6001 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     6061 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     6121 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     6181 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     6241 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     6301 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     6361 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgattg tcgacttaac
     6421 gcgtcaatta agcttgtgcc ccagtttgct agggaggtcg cagtatctgg ccactgccac
     6481 ctcgtgctgc tcgacgtagg tctcgttgtt ggcctccttg attctttcca gtctgtagtc
     6541 cacatagtag acgccaggca tcttgaggtt cttagcgggt ttcttggatc tatatgtggt
     6601 cttggcgttt gcgatcagat ggctcccgcc cacgagcttc agggccatgt cgtttctgcc
     6661 ttccaggccg ccgtcagcgg ggtacagagt ttcggtgaag gcctcccagc cgagtgtttt
     6721 cttctgcatc acagggccgt tggatgtgaa gttcacccct ctgatcttga cgttgtagat
     6781 gaggcagccg tcctggaggc tggtgtcctg ggtagcggtc agcacgcccc cgtcttcgta
     6841 tgtggtgact ctctcccatg tgaagccctc agggaaggac tgcttgaaga agtcggggat
     6901 gccctgggtg tggttgatga aggtcttgct gccgtagagg aagctagtag ccaggatgtc
     6961 gaaggcgaag gggagagggc cgccctcgac caccttgatt ctcatggtct gggtgccctc
     7021 gtagggcttg ccttcgccct cggatgtgca cttgaagtga tggttgtcca cggtgccctc
     7081 catgtacagc ttcatgtgca tgttctcctt aatcagctcg ctcataggac cagggttttc
     7141 ctcgacatca ccggcctgct tgagtagtga gaaattcgtg gcaccagatc ctctagactg
     7201 cagatccttt gccctaggac gagtgctggg gcgtctgttt ccactatcgg cgagcacttc
     7261 cacacagcca tcggtccaga cggcagcgct tctgcgggcg atttgtgtac gcccgacagt
     7321 cccggctccg gaccggacga ttgcgtcgca gcgaccctga gcccaagctg catcatcgaa
     7381 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tataagcccg
     7441 gagccgagga gatcctgcaa gctcgggatg cctccgctcg aagtagcggg tctgctgctc
     7501 catacaagcc aaccagggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
     7561 gaacatggcc tcgctccagt caatgacggc tgttatgcgg ccattgtcgg tcaggacatt
     7621 gttggagccg aaatcggcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
     7681 catcagctca tcgagagcct gggcgacgga ggcactgacg gtgtcgtcca tcacagtttg
     7741 ccagtgatac acatggggat cagcaatagc gcagatgaaa tcgcgccatg tagtgtattg
     7801 accgattcct tgaggtccga atgggccgaa cccgctggtc tggctaagat cggcggcagc
     7861 gatggcatcc atggcctcgg cgacgggctg gagaacagcg ggcagttcgg tttcaggcag
     7921 gtcttgcaag gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
     7981 ctccccaatg tcaagcactt cgggaatagg gagagcggcg gatgcaaagt gccgataaac
     8041 atagcgatct ttgtagaaac catcggcgca gctattcacc cgcaggacat atccacgccc
     8101 tcccacatcg aagctgaaag cccgagattc ttcgccctcg gagagctgca tcaggtcgga
     8161 tacactgtca aacttttcga tcagaaactt ctcgacagag gtagcggtga gttcaggctt
     8221 tttgaattcc atggtggccg tacgtcacga cacctgaaat ggaagaaaaa aactttgaac
     8281 cactgtctga ggcttgagaa tgaaccaaga tccaaactca aaaagggcaa attccaagga
     8341 gaattacatc aagtgccaag ctggcctaac ttcagtctcc acccactcag tgtggggaaa
     8401 ctccatcgca taaaacccct ccccccaacc taaagacgac gtactccaaa agctcgagaa
     8461 ctaatcgagg tgcctggacg gcgcccggta ctccgtggag tcacatgaag cgacggctga
     8521 ggacggaaag gcccttttcc tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg
     8581 cgtcctccat tttgagctcc ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc
     8641 aactggtgcc gaccgggcca gccttgccgc ccagggcggg gcgatacacg gcggcgcgag
     8701 gccaggcacc agagcaggcc ggccagcttg agactacccc cgtccgattc tcggtggccg
     8761 cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc
     8821 cgcggcccca aaaaccgaaa taccagtgtg cagatcttgg cccgcattta caagactatc
     8881 ttgccagaaa aaaagcgtcg cagcaggtca tcaaaaattt taaatggcta gagacttatc
     8941 gaaagcagcg agacaggcgc gaaggtgcca ccagattcgc acgcggcggc cccagcgccc
     9001 aggccaggcc tcaactcaag cacgaggcga aggggctcct taagcgcaag gcctcgaact
     9061 ctcccaccca cttccaaccc gaagctcggg atcaagaatc acgtactgca gccaggtgga
     9121 agtaattcaa ggcacgcaag ggccataacc cgtaaagagg ccaggcccgc gggaaccaca
     9181 cacggcactt acctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
     9241 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
     9301 atcactttcc cagtttaccc cgcgccacct tctctaggca ccggatcaat tgccgacccc
     9361 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga
     9421 gccaattccc actcctttca agacctagaa ggtccattag ctgcaaagat tcctctctgt
     9481 ttaaaacttt atccatcttt gcacccgggc cccctcgagc cggcgccaaa gtggatctct
     9541 gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt
     9601 agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat
     9661 cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtctcaaga tctagttacg
     9721 ccaagcttaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
     9781 attttaactt gctatttcta gctctaaaac agagacgtac aaaaaagagc aagaagctaa
     9841 aaaagattta aaaattattt ttagcgcagt taatggaaca ggaactaaat ttaccccaaa
     9901 aatattacgt gaatcaggat ataacgttat tgaggttgaa gagcatgcat ttgaagatga
     9961 aacatttaaa aatgttgtaa atccaaatcc agaatttgat cctgcatgaa aaataccgct
    10021 tgaatatggt attaaacatg atgcagatat tattattatg aatgacccag atgctgacag
    10081 atttggaatg gcaataaaac atgatggtca ttttgtaaga ttagatggaa atcaaacagg
    10141 accaatttta attgattgaa aattatcaaa tctaaaacgc ttaaatagca ttccaaaaaa
    10201 tccggctcta tattcaagtt ttgtaacaag tgatttgggt gatagaatcg ctcatgaaaa
    10261 atatggagtt aatattgtaa aaactttaac tggatttaaa tgaatgggta gagaaattgc
    10321 taaagaagaa gataacggat taaattttgt ttttgcttat gaagaaagtt atggatatgt
    10381 aattgatgac tcagctagag ataaagatgg aatacaagct tctatattaa tagcagaggc
    10441 tgcttgattt tataaaaaac aaaataaaac attagtagac tatttagaag atttatttaa
    10501 agaaatgggt gcatattaca ctttcacttt aaacttgaat tttaaaccag aagaaaagaa
    10561 attaaaaatt gaaccattaa tgaaatcatt gagagcaaca cccttaactc aaattgctgg
    10621 acttaaagtt gttaatgttg aagactacat cgatggaatg tataatatgc caggacaaga
    10681 cttactaaaa ttttatttag aagataagtc atgatttgct gttcgcccaa gtggaactga
    10741 acctaaacta aaaatttatt ttataggtgt tggtgaatct gttcaaaacg ctaaagttaa
    10801 agtagacgaa attattaaag aattaaaatt aaaaatgaat atataggaga aaaaatgaaa
    10861 ctaaacaaat atatagatca cacattatta aaacaagatg ctacgaaagc tgaaattaaa
    10921 caattatgtg atgaagcaat tgaatttgat tttgcaacag tttgtgttaa ttcatattga
    10981 acaagctatt gtaaagaatt attaaaaggc acaaatgtag gaataacaaa tgttgtaggt
    11041 tttcctctag gtgcatgcac aacagctaca aaagcattcg aagtttctga agcaattaaa
    11101 gatggtgcaa cagaaattga tatggtatta aatattggtg cattaaaaga caaaaattat
    11161 gaattagttt tagaagacat gaaagctgta aaaaaagcag ctggatcaca tgttgttaaa
    11221 tgtattatgg aaaattgttt attaacaaaa gaagaaatca tgaaagcttg tgaaatagct
    11281 gttgaagctg gattagaatt tgttaaaaca tcaacaggat tttcaaaatc aggtgcaaca
    11341 tttgaagatg ttaaactaat gaagtcagtt gttaaagaca atgctttagt t
//
