LOCUS       sequence-315769-        9562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty TET-On expression of CRISPRi with gSTOP gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9562)
  AUTHORS   Backstrom JR, Sheng J, Wang MC, Bernardo-Colon A, Rex TS
  TITLE     Optimization of S. aureus dCas9 and CRISPRi Elements for a Single 
            Adeno-Associated Virus that Targets an Endogenous Gene.
  JOURNAL   Mol Ther Methods Clin Dev. 2020 Sep 6;19:139-148. doi: 
            10.1016/j.omtm.2020.09.001. eCollection 2020 Dec 11.
  PUBMED    33024795
REFERENCE   2  (bases 1 to 9562)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2020-09-6"; volume: "19:139-148. doi"; 
            pages: " 10.1016/j.omtm.2020.09.001. eCollection 2020 Dec 11"
COMMENT     Sequence Label: (517) SB KRAB-SadCas9 U6-gStop
FEATURES             Location/Qualifiers
     source          1..9562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        295..607
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    303..321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    339..357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    374..392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    410..428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    446..464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    481..499
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    517..535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     580..604
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             694..879
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             4132..4158
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     polyA_signal    4202..4323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4213..4232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4267..4286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4362..4610
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4362..4382
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4533..4552
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4631..4706
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     CDS             join(5347..5964,5965..5970,5971..6006,6007..6009,6010..6045,
                     6046..6048,6049..6084,6085..6090)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             6097..6153
                     /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             6148..6948
                     /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(6208..6227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6818..6837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7003..7020)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7009..7233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7559..7576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7730..8318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7810..7829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8489..9280,9281..9349))
                     /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     9112..9131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9350..9454)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agacgagttt
      301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
      481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
      541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta
      661 ccctcgaaag gcctctgaga attcgccacc atgcggacac tagtgacctt caaggatgta
      721 tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac
      781 agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag
      841 ccagatgtga tcctccggtt ggagaaggga gaagagcccg gcggttccgg cggagggtcc
      901 atgggcaaac ggaactacat cctggggctt gccattggga taaccagcgt tggctacgga
      961 attattgatt atgagacacg cgatgtgatt gacgccgggg ttaggctgtt caaagaggcc
     1021 aacgttgaaa acaacgaggg aagacggagt aagcgcggag caagaagact caagcgcaga
     1081 cggagacatc ggattcagag ggtgaaaaag ctgctcttcg attacaatct cctgaccgat
     1141 catagtgagc tgagcggaat caacccctac gaggcgcgag tgaaagggct ttcccagaag
     1201 ctgtccgaag aggagttctc cgccgcgttg ctgcacctgg ccaaacggag gggggttcac
     1261 aatgtaaacg aagtggagga ggacacgggc aatgaactta gtacgaaaga acagatcagt
     1321 aggaactcta aggctctcga agagaaatac gtcgctgagt tgcagcttga gagactgaaa
     1381 aaagacggcg aagtacgcgg atctattaat aggttcaaga cttcagatta cgtaaaggaa
     1441 gccaagcagc tcctgaaagt acagaaagcg taccatcagc tcgatcagag cttcatcgat
     1501 acctacatag atttgctgga gacacggagg acatactacg agggcccagg ggaaggatct
     1561 ccttttgggt ggaaggacat caaggaatgg tacgagatgc ttatgggaca ttgtacatat
     1621 tttccggagg agctcaggag cgtcaagtac gcctacaatg ccgacctgta caatgccctc
     1681 aatgacctca ataacctcgt gattaccagg gacgagaacg agaagctgga gtactatgaa
     1741 aagttccaga ttatcgagaa tgtgtttaag cagaagaaga agccgacact taagcagatt
     1801 gcaaaggaaa tcctcgtgaa tgaggaagat atcaagggat acagagtgac aagtacaggc
     1861 aagcccgagt tcacaaatct gaaggtgtac cacgatatta aggacataac cgcacgaaag
     1921 gagataatcg aaaacgctga gctcctcgat cagatcgcaa aaattcttac catctaccag
     1981 tctagtgagg acattcagga ggaactgact aatctgaaca gtgagctcac ccaagaggaa
     2041 attgagcaga tttcaaacct gaaaggctac accgggacgc acaatctgag cctcaaagca
     2101 atcaacctca ttctggatga actttggcac acaaatgaca accaaattgc catattcaac
     2161 cgcctgaaac tggtgccaaa aaaagtggat ctgtcacagc aaaaggaaat ccctacaacc
     2221 ttggttgacg attttattct gtcccccgtt gtcaagcgga gcttcatcca gtcaatcaag
     2281 gtgatcaatg ccatcattaa aaaatacgga ttgccaaacg atataattat cgagcttgca
     2341 cgagagaaga actcaaagga cgcccagaag atgattaacg aaatgcagaa gcgcaaccgc
     2401 cagacaaacg aacgcataga ggaaattata agaacaaccg gcaaagagaa tgccaagtat
     2461 ctgatcgaga aaatcaagct gcacgacatg caagaaggca agtgcctgta ctctctggaa
     2521 gctatcccac tcgaagatct gctgaataat ccattcaatt acgaggtgga cgccatcatc
     2581 cctagatccg taagctttga caattccttc aataacaaag ttctggttaa acaggaggaa
     2641 aattctaaaa aagggaaccg gaccccgttc cagtacctga gctccagtga cagcaagatt
     2701 agctacgaga cttttaagaa acatattctg aatctggcca aaggcaaagg caggatcagc
     2761 aagaccaaga aggagtacct cctcgaagaa cgcgacatta acagatttag tgtgcagaaa
     2821 gatttcatca accgaaacct tgtcgatact cggtacgcca cgagaggcct gatgaatctc
     2881 ctcaggagct acttccgcgt caataatctg gacgttaaag tcaagagcat aaatggggga
     2941 ttcaccagct ttctgaggag aaagtggaag tttaagaagg aacgaaacaa aggatacaag
     3001 caccatgctg aggatgcttt gatcatcgct aacgcggact ttatctttaa ggaatggaaa
     3061 aagctggata aggcaaagaa agtgatggaa aaccagatgt tcgaggagaa gcaggcagag
     3121 tcaatgcctg agatcgagac agagcaggaa tacaaggaaa ttttcatcac ccctcatcag
     3181 attaaacaca taaaggactt caaagactat aaatactctc atagggtgga caaaaaaccc
     3241 aatcgcgagc tcattaatga caccctgtac tcaacacgga aggatgataa aggtaatacc
     3301 ttgattgtga ataatcttaa tggattgtat gacaaagata acgacaagct caagaagctg
     3361 atcaacaagt ctccagagaa gctccttatg tatcaccacg acccacagac ttatcagaaa
     3421 ttgaaactga tcatggagca atacggggat gagaagaacc cactctacaa atattatgag
     3481 gaaacaggta attacctgac caagtactcc aagaaggata acggaccagt gatcaaaaag
     3541 ataaagtact atggcaacaa acttaatgcg catttggaca taactgacga ttaccccaat
     3601 tctcgaaaca aggttgtgaa gctctccctg aagccttata gatttgacgt gtacctggat
     3661 aatggggttt ataaattcgt caccgtgaaa aatctggacg tgatcaaaaa ggagaactat
     3721 tatgaagtaa actcaaagtg ctatgaggag gcgaagaagc tgaagaagat ctccaatcag
     3781 gccgagttca tcgcttcctt ctataataac gatctcatca agatcaatgg agagctttat
     3841 cgcgtcattg gtgtgaacaa tgacttgctg aacaggatcg aagtcaatat gatagacatt
     3901 acctaccggg agtatctcga aaacatgaat gataaacggc cgcctcgcat catcaagaca
     3961 atcgcatcta aaactcagtc aataaaaaag tactctaccg atatcctggg gaatctctat
     4021 gaagtgaagt caaagaagca cccacaaatc attaaaaaag gtggatccgc agaagcagca
     4081 gcaaaggaag cagcagcaaa ggaagcagca gcaaaggcag tcgacaccgg tccagcagct
     4141 aagagagtta aactagatta gcggccgcat cgtcaggcca agcttccatc gatagacatg
     4201 ataagataca ttgatgagtt tggacaaacc acaacaagaa tgcagtgaaa aaaatgcttt
     4261 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4321 gttaacaaca acaattgcat tcattttatg tttcaggtac cgagggccta tttcccatga
     4381 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
     4441 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     4501 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     4561 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagaccaag
     4621 gcagtttttt gttttagtac tctggaaaca gaatctacta aaacaaggca aaatgccgtg
     4681 tttatctcgt caacttgttg gcgagatttt tgcggcctag gcgagaccct gtctcacaaa
     4741 ataaagtaag cccggactga gtgcggaaag gcgggcctgg cgggtctggt ctccccatgc
     4801 gggccaccag aggccctgca gccttcagtc gcttgaaggg gtaatggcgc ttccactcac
     4861 aaacatggcg gacagagcgt gtgaacgaga tgaacagccc ctcaaaaata tggccgccga
     4921 ggctggacgg ccgtgcccca gcagcaccgc ctccgcgccc cacgtgatct ctcgccgggc
     4981 acagcgctga ccgcggaggt ccaaccggaa gaatgtccgg attggacatt cggaagaggg
     5041 cccgccttcc ctggggaatc tctgcgcacg cgcagaacgc ttcgaccaat gaaaacacag
     5101 gaagccgtcc gcgcaaccgc gttgcgtcac ttctgccgcc cctgtttcaa ggtatatagc
     5161 cgtagacgga acttcgcctt tctctcggcc ttagcgccat ttttttgggt gagtgttttt
     5221 tggttcctgc gttgggattc cgtgtacaat ccatagacat ctgacctcgg cacttagcat
     5281 catcacagca aactaactgt agcctttctc tctttccctg tagaaacctc tgcacctgag
     5341 gccaccatgt ctagactgga caagagcaaa gtcataaacg gcgctctgga attactcaat
     5401 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
     5461 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     5521 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
     5581 ctgcggaaca acgccaagtc attccgctgt gctctcctct cacatcgcga cggggctaaa
     5641 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     5701 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     5761 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     5821 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
     5881 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     5941 aaacagctaa agtgcgaaag cggcgggccg gccgacgccc ttgacgattt tgacttagac
     6001 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     6061 gacgattttg accttgacat gctccccggg tccggcgcta ctaacttcag cctgctgaag
     6121 caggctggtg acgtcgagga gaatcctggt cccatgattg aacaagatgg attgcacgca
     6181 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6241 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6301 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6361 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6421 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6481 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6541 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6601 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6661 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccacggt
     6721 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6781 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6841 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6901 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaggcctg
     6961 tcgtgaagct tggggatcaa ttctctagag ctcgctgatc agcctcgact gtgccttcta
     7021 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     7081 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     7141 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     7201 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     7261 cctactagct actcgggacc ccttaccgaa acatcgccgc attctgcaga ggagtcgagt
     7321 gtatgtaaac ttctgaccca ctgggaatgt gatgaaagaa ataaaagctg aaatgaatca
     7381 ttctctctac tattattctg atatttcaca ttcttaaaat aaagtggtga tcctaactga
     7441 cctaagacag ggaattttta ctaggattaa atgtcaggaa ttgtgaaaaa gtgagtttaa
     7501 atgtatttgg ctaaggtgta tgtaaacttc cgacttcaac tgtataggga tccgcttcct
     7561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8401 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8461 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8521 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8581 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8641 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8701 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8761 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8821 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8881 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8941 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9001 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9061 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9121 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9181 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9241 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9301 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9361 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9421 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9481 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
     9541 gcgttaatat tttgttaaaa tt
//
