LOCUS       sequence-321587-        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        284..596
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    292..310
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    328..346
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    399..417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    435..453
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    470..488
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    506..524
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     569..593
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             683..868
                     /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             4121..4147
                     /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    4191..4312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4202..4221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4256..4275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4351..4599
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4351..4371
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4522..4541
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4620..4695
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     CDS             join(5336..5953,5954..5959,5960..5995,5996..5998,5999..6034,
                     6035..6037,6038..6073,6074..6079)
                     /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             6086..6142
                     /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             6137..6937
                     /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(6197..6216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6807..6826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6992..7009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6998..7222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7548..7565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7719..8307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7799..7818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8478..9269,9270..9338))
                     /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     9101..9120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9339..9443)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagttgaagt cggaagttta catacactta agttggagtc attaaaactc gtttttcaac
       61 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
      121 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
      181 attcactgta tcacaattcc agtgggtcag aagtttacat acactaagtt cgactcctct
      241 gcagaatgcg gcgatgtttc ggtaaggggt ccgctatcta gacgagttta ctccctatca
      301 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      421 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
      481 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgattagg
      541 cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga
      601 gccaattcca actcttttgc cttataccta ctttccgtac cacttcctac cctcgaaagg
      661 cctctgagaa ttcgccacca tgcggacact agtgaccttc aaggatgtat ttgtggactt
      721 caccagggag gagtggaagc tgctggacac tgctcagcag atcgtgtaca gaaatgtgat
      781 gctggagaac tataagaacc tggtttcctt gggttatcag cttactaagc cagatgtgat
      841 cctccggttg gagaagggag aagagcccgg cggttccggc ggagggtcca tgggcaaacg
      901 gaactacatc ctggggcttg ccattgggat aaccagcgtt ggctacggaa ttattgatta
      961 tgagacacgc gatgtgattg acgccggggt taggctgttc aaagaggcca acgttgaaaa
     1021 caacgaggga agacggagta agcgcggagc aagaagactc aagcgcagac ggagacatcg
     1081 gattcagagg gtgaaaaagc tgctcttcga ttacaatctc ctgaccgatc atagtgagct
     1141 gagcggaatc aacccctacg aggcgcgagt gaaagggctt tcccagaagc tgtccgaaga
     1201 ggagttctcc gccgcgttgc tgcacctggc caaacggagg ggggttcaca atgtaaacga
     1261 agtggaggag gacacgggca atgaacttag tacgaaagaa cagatcagta ggaactctaa
     1321 ggctctcgaa gagaaatacg tcgctgagtt gcagcttgag agactgaaaa aagacggcga
     1381 agtacgcgga tctattaata ggttcaagac ttcagattac gtaaaggaag ccaagcagct
     1441 cctgaaagta cagaaagcgt accatcagct cgatcagagc ttcatcgata cctacataga
     1501 tttgctggag acacggagga catactacga gggcccaggg gaaggatctc cttttgggtg
     1561 gaaggacatc aaggaatggt acgagatgct tatgggacat tgtacatatt ttccggagga
     1621 gctcaggagc gtcaagtacg cctacaatgc cgacctgtac aatgccctca atgacctcaa
     1681 taacctcgtg attaccaggg acgagaacga gaagctggag tactatgaaa agttccagat
     1741 tatcgagaat gtgtttaagc agaagaagaa gccgacactt aagcagattg caaaggaaat
     1801 cctcgtgaat gaggaagata tcaagggata cagagtgaca agtacaggca agcccgagtt
     1861 cacaaatctg aaggtgtacc acgatattaa ggacataacc gcacgaaagg agataatcga
     1921 aaacgctgag ctcctcgatc agatcgcaaa aattcttacc atctaccagt ctagtgagga
     1981 cattcaggag gaactgacta atctgaacag tgagctcacc caagaggaaa ttgagcagat
     2041 ttcaaacctg aaaggctaca ccgggacgca caatctgagc ctcaaagcaa tcaacctcat
     2101 tctggatgaa ctttggcaca caaatgacaa ccaaattgcc atattcaacc gcctgaaact
     2161 ggtgccaaaa aaagtggatc tgtcacagca aaaggaaatc cctacaacct tggttgacga
     2221 ttttattctg tcccccgttg tcaagcggag cttcatccag tcaatcaagg tgatcaatgc
     2281 catcattaaa aaatacggat tgccaaacga tataattatc gagcttgcac gagagaagaa
     2341 ctcaaaggac gcccagaaga tgattaacga aatgcagaag cgcaaccgcc agacaaacga
     2401 acgcatagag gaaattataa gaacaaccgg caaagagaat gccaagtatc tgatcgagaa
     2461 aatcaagctg cacgacatgc aagaaggcaa gtgcctgtac tctctggaag ctatcccact
     2521 cgaagatctg ctgaataatc cattcaatta cgaggtggac gccatcatcc ctagatccgt
     2581 aagctttgac aattccttca ataacaaagt tctggttaaa caggaggaaa attctaaaaa
     2641 agggaaccgg accccgttcc agtacctgag ctccagtgac agcaagatta gctacgagac
     2701 ttttaagaaa catattctga atctggccaa aggcaaaggc aggatcagca agaccaagaa
     2761 ggagtacctc ctcgaagaac gcgacattaa cagatttagt gtgcagaaag atttcatcaa
     2821 ccgaaacctt gtcgatactc ggtacgccac gagaggcctg atgaatctcc tcaggagcta
     2881 cttccgcgtc aataatctgg acgttaaagt caagagcata aatgggggat tcaccagctt
     2941 tctgaggaga aagtggaagt ttaagaagga acgaaacaaa ggatacaagc accatgctga
     3001 ggatgctttg atcatcgcta acgcggactt tatctttaag gaatggaaaa agctggataa
     3061 ggcaaagaaa gtgatggaaa accagatgtt cgaggagaag caggcagagt caatgcctga
     3121 gatcgagaca gagcaggaat acaaggaaat tttcatcacc cctcatcaga ttaaacacat
     3181 aaaggacttc aaagactata aatactctca tagggtggac aaaaaaccca atcgcgagct
     3241 cattaatgac accctgtact caacacggaa ggatgataaa ggtaatacct tgattgtgaa
     3301 taatcttaat ggattgtatg acaaagataa cgacaagctc aagaagctga tcaacaagtc
     3361 tccagagaag ctccttatgt atcaccacga cccacagact tatcagaaat tgaaactgat
     3421 catggagcaa tacggggatg agaagaaccc actctacaaa tattatgagg aaacaggtaa
     3481 ttacctgacc aagtactcca agaaggataa cggaccagtg atcaaaaaga taaagtacta
     3541 tggcaacaaa cttaatgcgc atttggacat aactgacgat taccccaatt ctcgaaacaa
     3601 ggttgtgaag ctctccctga agccttatag atttgacgtg tacctggata atggggttta
     3661 taaattcgtc accgtgaaaa atctggacgt gatcaaaaag gagaactatt atgaagtaaa
     3721 ctcaaagtgc tatgaggagg cgaagaagct gaagaagatc tccaatcagg ccgagttcat
     3781 cgcttccttc tataataacg atctcatcaa gatcaatgga gagctttatc gcgtcattgg
     3841 tgtgaacaat gacttgctga acaggatcga agtcaatatg atagacatta cctaccggga
     3901 gtatctcgaa aacatgaatg ataaacggcc gcctcgcatc atcaagacaa tcgcatctaa
     3961 aactcagtca ataaaaaagt actctaccga tatcctgggg aatctctatg aagtgaagtc
     4021 aaagaagcac ccacaaatca ttaaaaaagg tggatccgca gaagcagcag caaaggaagc
     4081 agcagcaaag gaagcagcag caaaggcagt cgacaccggt ccagcagcta agagagttaa
     4141 actagattag cggccgcatc gtcaggccaa gcttccatcg atagacatga taagatacat
     4201 tgatgagttt ggacaaacca caacaagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4321 caattgcatt cattttatgt ttcaggtacc gagggcctat ttcccatgat tccttcatat
     4381 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     4441 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     4501 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     4561 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagaccaagg cagttttttg
     4621 ttttagtact ctggaaacag aatctactaa aacaaggcaa aatgccgtgt ttatctcgtc
     4681 aacttgttgg cgagattttt gcggcctagg cgagaccctg tctcacaaaa taaagtaagc
     4741 ccggactgag tgcggaaagg cgggcctggc gggtctggtc tccccatgcg ggccaccaga
     4801 ggccctgcag ccttcagtcg cttgaagggg taatggcgct tccactcaca aacatggcgg
     4861 acagagcgtg tgaacgagat gaacagcccc tcaaaaatat ggccgccgag gctggacggc
     4921 cgtgccccag cagcaccgcc tccgcgcccc acgtgatctc tcgccgggca cagcgctgac
     4981 cgcggaggtc caaccggaag aatgtccgga ttggacattc ggaagagggc ccgccttccc
     5041 tggggaatct ctgcgcacgc gcagaacgct tcgaccaatg aaaacacagg aagccgtccg
     5101 cgcaaccgcg ttgcgtcact tctgccgccc ctgtttcaag gtatatagcc gtagacggaa
     5161 cttcgccttt ctctcggcct tagcgccatt tttttgggtg agtgtttttt ggttcctgcg
     5221 ttgggattcc gtgtacaatc catagacatc tgacctcggc acttagcatc atcacagcaa
     5281 actaactgta gcctttctct ctttccctgt agaaacctct gcacctgagg ccaccatgtc
     5341 tagactggac aagagcaaag tcataaacgg cgctctggaa ttactcaatg gagtcggtat
     5401 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     5461 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     5521 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     5581 cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     5641 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     5701 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     5761 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     5821 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     5881 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     5941 gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca tgctcccagc
     6001 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     6061 ccttgacatg ctccccgggt ccggcgctac taacttcagc ctgctgaagc aggctggtga
     6121 cgtcgaggag aatcctggtc ccatgattga acaagatgga ttgcacgcag gttctccggc
     6181 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6241 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6301 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     6361 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6421 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6481 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6541 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6601 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6661 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccacggtg atgcctgctt
     6721 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6781 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6841 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6901 catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaggcctgt cgtgaagctt
     6961 ggggatcaat tctctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     7021 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     7081 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7141 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7201 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggc ctactagcta
     7261 ctcgggaccc cttaccgaaa catcgccgca ttctgcagag gagtcgagtg tatgtaaact
     7321 tctgacccac tgggaatgtg atgaaagaaa taaaagctga aatgaatcat tctctctact
     7381 attattctga tatttcacat tcttaaaata aagtggtgat cctaactgac ctaagacagg
     7441 gaatttttac taggattaaa tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc
     7501 taaggtgtat gtaaacttcc gacttcaact gtatagggat ccgcttcctc gctcactgac
     7561 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7621 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7681 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7741 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7801 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7861 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7921 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7981 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8041 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8101 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8161 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8221 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8281 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8341 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8401 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8521 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8881 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga
     9481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     9541 ttgttaaaat tggatcccta ta
//
