LOCUS       sequence-395062-       10556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tamoxifen-inducible expression of sgRNA control targeting intergenic
            region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10556)
  AUTHORS   So J, Mabe NW, Englinger B, Chow KH, Moyer SM, Yerrum S, Trissal MC,
            Marques JG, Kwon JJ, Shim B, Pal S, Panditharatna E, Quinn T, 
            Schaefer DA, Jeong D, Mayhew DL, Hwang J, Beroukhim R, Ligon KL, 
            Stegmaier K, Filbin MG, Hahn WC
  TITLE     VRK1 as a synthetic lethal target in VRK2 promoter-methylated 
            cancers of the nervous system.
  JOURNAL   JCI Insight. 2022 Oct 10;7(19):e158755. doi: 
            10.1172/jci.insight.158755.
  PUBMED    36040810
REFERENCE   2  (bases 1 to 10556)
  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"; journalName: "JCI 
            Insight."; date: "2022-10-10"; volume: "7(19):e158755. doi"; pages: 
            " 10.1172/jci.insight.158755"
COMMENT     Sequence Label: pLenti-SwitchON-sgCh2-2
FEATURES             Location/Qualifiers
     source          1..10556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        242..621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        623..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     776..796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             839..1019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1066..1191
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2655..2673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2691..2709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2726..2744
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2762..2780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2798..2816
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2833..2851
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2869..2887
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3217..3465
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3217..3237
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3388..3407
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(3504..3537)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3577..3610)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3926..3949)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4276..4295
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4374..4430
                     /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             4425..5225
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4485..4504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5095..5114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    5248..5836
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5301..5321)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5719..5730)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5839..5855)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5840..5856)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6361..6541
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6567..6584)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6573..6797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6843..7271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6930..6949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7140..7161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7280..7300)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7285..7614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7465..7600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7527..7546
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7662..7709
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7728..8102
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8232..8353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8269..8288)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8323..8342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8402..8418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8402..8418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8415..8437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8426..8442
                     /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(8450..8456,8457..8474,8475..8480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8495..8516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8633..8650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8804..9392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8884..8903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9563..10354,10355..10423))
                     /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     10186..10205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10424..10528)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgcgcgt
      241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
      841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     1021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     1081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     1141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     1201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     1261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     1321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     1381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     1441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     1501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 ggctagcctc taggcgccgg aattgaagat ctccgcggtt aattaaaatg atacggaacg
     2641 tatgaagagt ttactcccta tcagtgatag agaacgtatg ymgastttac tccctatcag
     2701 tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
     2761 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
     2821 atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
     2881 atagagaacg tatgtcgagg taggtgtgta cgggccccag aaacattgtg acttcgtggc
     2941 agagcccatg ggggagaagc cagtggggag cctggctggg attggtgaag tcctgggcaa
     3001 gaagctggag gaaaggggtt ttgacaaggc ctatgttgtc cttggccagt ttctggtgct
     3061 aaagaaagat gaagacctct tccgggaatg gctgaaagac acttgtggcg ccaacgccaa
     3121 gcagtcccgg gactgcttcg gatgccttcg agagtggtgc gacgccgacc accggatctg
     3181 tcttgatcta ggagctactc atcaacggtc acatgtgagg gcctatttcc catgattcct
     3241 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     3301 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     3361 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     3421 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgggtg tgcgtatgaa
     3481 gcagtggttt aagagctatg ctgataactt cgtatagcat acattatacg aagttatttt
     3541 ttttttttgc ggccgcccag aattctggcc tgcaggataa cttcgtatag catacattat
     3601 acgaagttat cagcatagca agtttaaata aggctagtcc gttatcaact tgaaaaagtg
     3661 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
     3721 ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac ccgttaagat
     3781 cggaagagca cacgtctgaa ctccagtcac aaaattatct cgtatgccgt cttatgcttg
     3841 ttaattaaca attgacgcgt aattctaccg gctagcgatc taattctacc gggtagggga
     3901 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     3961 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     4021 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     4081 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac ctctcactag
     4141 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     4201 ccaatagcag ctttgctcct tcgctttccg ggctcagggg cggggcgggc gcccgaaggt
     4261 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     4321 cttcctcatc tccgggcctt tcgacctgca gccaattgta caagggcagc ggcgccacca
     4381 acttcagcct gctgaagcag gccggcgacg tggaggagaa ccccggcccc atgattgaac
     4441 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4501 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4561 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4621 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4681 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4741 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4801 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4861 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4921 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4981 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5041 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5101 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5161 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5221 tctgaattcg atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga
     5281 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5341 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5401 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5461 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5521 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5581 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5641 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5701 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5761 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5821 tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc
     5881 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     5941 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     6001 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6061 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     6121 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
     6181 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
     6241 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
     6301 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
     6361 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6421 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6481 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6541 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     6601 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6661 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6721 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     6781 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     6841 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6901 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6961 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     7021 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     7081 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     7141 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     7201 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     7261 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     7321 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     7381 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7441 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7501 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     7561 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     7621 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     7681 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     7741 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     7801 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     7861 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     7921 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     7981 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     8041 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     8101 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     8161 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     8221 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     8281 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8341 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     8401 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     8461 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     8521 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8581 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8641 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8701 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8761 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8821 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8881 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8941 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     9001 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     9061 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     9121 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     9181 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     9241 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     9301 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     9361 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     9421 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9481 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9541 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9601 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9661 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9721 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9781 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9841 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9901 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9961 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    10021 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    10081 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    10141 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    10201 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    10261 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    10321 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10381 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    10441 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    10501 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
