LOCUS       sequence-191025-       11409 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 eGFP
            from EF-1a promoter. 3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11409)
  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 11409)
  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-eGFP
FEATURES             Location/Qualifiers
     source          1..11409
                     /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             join(2328..2330,2331..2333,2334..3047)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2328..2330,2331..2333,2334..3044)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2373..2394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2634..2653)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2981..3002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3062..3650
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3115..3135)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3533..3544)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3722..3955
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4033..4154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4070..4089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4124..4143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4194..4329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4349..4368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4350..4368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4378..4395)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4378..4394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4387..4409)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4536..4991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4623..4642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4833..4854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5017..5121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5122..5190,5191..5982)
                     /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(5340..5359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6153..6741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6642..6661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6895..6912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7029..7050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7065..7070,7071..7088,7089..7095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7103..7119
                     /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     7108..7130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7127..7143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7127..7143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7162..7182
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7164..7182
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7210..7436
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7437..7617
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7664..7789
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8282..8515
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8700..8744
                     /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        8922..9170
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8922..8942
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9093..9112
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11052..11127
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    11183..11300
                     /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 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgacgc gttaagtcga
     3061 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3121 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3181 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3241 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3301 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3361 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3421 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3481 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3541 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3601 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     3661 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     3721 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     3781 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     3841 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     3901 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     3961 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     4021 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4081 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4141 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     4201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4261 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4321 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     4381 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4441 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4501 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4561 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4621 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4681 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4741 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4801 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4861 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4921 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4981 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5821 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6301 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6361 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6421 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6481 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6541 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6601 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6661 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6721 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6781 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6841 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6901 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6961 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7021 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7141 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     7201 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     7261 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     7321 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     7381 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     7441 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7501 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7561 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     7621 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     7681 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     7741 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     7801 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     7861 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     7921 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     7981 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     8041 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     8101 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     8161 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     8221 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     8281 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     8341 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     8401 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     8461 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     8521 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     8581 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     8641 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     8701 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     8761 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     8821 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     8881 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
     8941 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
     9001 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     9061 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     9121 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt
     9181 gtaaatgagc acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc
     9241 aaccaaaatc ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt
     9301 tgcaaacttt taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg
     9361 ttttcttcac ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct
     9421 tcaatcattt tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca
     9481 ttgtctttaa caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa
     9541 aatcctgttg atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc
     9601 atgatttctt cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca
     9661 gctgcttttt ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat
     9721 gcaccaatat ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact
     9781 tcgaatgctt ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt
     9841 cctacatttg tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa
     9901 actgttgcaa aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta
     9961 gcatcttgtt ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata
    10021 tattcatttt taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa
    10081 cagattcacc aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa
    10141 cagcaaatca tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat
    10201 acattccatc gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg
    10261 gtgttgctct caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa
    10321 aattcaagtt taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat
    10381 agtctactaa tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag
    10441 aagcttgtat tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt
    10501 cataagcaaa aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc
    10561 atttaaatcc agttaaagtt tttacaatat taactccata tttttcatga gcgattctat
    10621 cacccaaatc acttgttaca aaacttgaat atagagccgg attttttgga atgctattta
    10681 agcgttttag atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta
    10741 atcttacaaa atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat
    10801 tcataataat aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag
    10861 gatcaaattc tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct
    10921 cttcaacctc aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc
    10981 ctgttccatt aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt
    11041 tgtacgtctc tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
    11101 cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag atcttgagac
    11161 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
    11221 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
    11281 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
    11341 ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca
    11401 gctaatgga
//
