LOCUS       sequence-191027-       11394 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 
            dTomato from EF-1a promoter. 3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11394)
  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 11394)
  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-dTomato
FEATURES             Location/Qualifiers
     source          1..11394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..1208
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          261..1199
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1156..1176
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1215..1224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1230..2246
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2271..2327
                     /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             2328..3029
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2461..2479)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2729..2748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2923..2946
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3047..3635
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3100..3120)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3518..3529)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3707..3940
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4018..4139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4055..4074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4109..4128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4179..4314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4334..4353)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4335..4353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4363..4380)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4363..4379)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4372..4394)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4521..4976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4608..4627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4818..4839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5002..5106
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5107..5175,5176..5967)
                     /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(5325..5344)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6138..6726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6627..6646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6880..6897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7014..7035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7050..7055,7056..7073,7074..7080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7088..7104
                     /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     7093..7115
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7112..7128
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7112..7128
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7147..7167
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7149..7167
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7195..7421
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7422..7602
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7649..7774
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8267..8500
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8685..8729
                     /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"
     promoter        8907..9155
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8907..8927
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9078..9097
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11037..11112
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    11168..11285
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
       61 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga
      121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      241 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
      301 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
      361 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
      421 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
      481 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
      541 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
      601 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
      661 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
      721 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
      781 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
      841 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
      901 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
      961 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     1021 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1081 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1141 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1201 tgtcgtgacg tacggccacc atggaattca aaaagcctga actcaccgct acctctgtcg
     1261 agaagtttct gatcgaaaag tttgacagtg tatccgacct gatgcagctc tccgagggcg
     1321 aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg cgggtgaata
     1381 gctgcgccga tggtttctac aaagatcgct atgtttatcg gcactttgca tccgccgctc
     1441 tccctattcc cgaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct
     1501 cccgccgtgc acagggtgtc accttgcaag acctgcctga aaccgaactg cccgctgttc
     1561 tccagcccgt cgccgaggcc atggatgcca tcgctgccgc cgatcttagc cagaccagcg
     1621 ggttcggccc attcggacct caaggaatcg gtcaatacac tacatggcgc gatttcatct
     1681 gcgctattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     1741 cctccgtcgc ccaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     1801 ggcacctcgt gcacgccgat ttcggctcca acaatgtcct gaccgacaat ggccgcataa
     1861 cagccgtcat tgactggagc gaggccatgt tcggggattc ccaatacgag gtcgccaaca
     1921 tcttcttctg gaggccctgg ttggcttgta tggagcagca gacccgctac ttcgagcgga
     1981 ggcatcccga gcttgcagga tctcctcggc tccgggctta tatgctccgc attggtcttg
     2041 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gctcagggtc
     2101 gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     2161 gaagcgctgc cgtctggacc gatggctgtg tggaagtgct cgccgatagt ggaaacagac
     2221 gccccagcac tcgtcctagg gcaaaggatc tgcagtctag aggatctggt gccacgaatt
     2281 tctcactact caagcaggcc ggtgatgtcg aggaaaaccc tggtcctatg gtgagcaagg
     2341 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     2401 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2461 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     2521 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     2581 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     2641 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     2701 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2761 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     2821 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     2881 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     2941 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     3001 tcctgtacgg catggacgag ctgtacaagt gacgcgttaa gtcgacaatc aacctctgga
     3061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3121 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3241 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3301 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3361 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3421 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3481 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3541 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3601 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
     3661 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     3721 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     3781 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     3841 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     3901 tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt
     3961 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     4021 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4081 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4141 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4201 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4261 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga
     4321 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     4381 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     4441 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4501 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4861 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4921 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     4981 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5041 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5101 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5161 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5221 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5281 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5341 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5401 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5461 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5521 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5581 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5641 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5701 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5761 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5821 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5881 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5941 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6001 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6061 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6781 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6841 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6901 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6961 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7021 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7081 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7141 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt
     7201 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
     7261 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
     7321 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
     7381 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
     7441 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7501 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7561 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     7621 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     7681 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     7741 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     7801 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     7861 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     7921 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     7981 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     8041 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     8101 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     8161 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     8221 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     8281 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     8341 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     8401 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     8461 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     8521 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     8581 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     8641 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     8701 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     8761 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     8821 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     8881 acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc
     8941 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
     9001 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     9061 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     9121 tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca
     9181 aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt
     9241 tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata
     9301 aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta
     9361 ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga
     9421 gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact
     9481 gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt
     9541 ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt
     9601 gttaataaac aattttccat aatacattta acaacatgtg atccagctgc tttttttaca
     9661 gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat
     9721 accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta
     9781 gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct
     9841 tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca
     9901 aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat
     9961 aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt
    10021 ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac
    10081 ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact
    10141 tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt
    10201 agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg
    10261 atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag
    10321 tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt
    10381 tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat
    10441 ctttatctct agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa
    10501 aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta
    10561 aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg
    10621 ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg
    10681 ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac
    10741 catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat
    10801 ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat
    10861 ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa
    10921 cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg
    10981 cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt
    11041 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
    11101 ccgagtcggt gcttttttaa gcttggcgta actagatctt gagacaaatg gcagtattca
    11161 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
    11221 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
    11281 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc
    11341 gggtgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tgga
//
