LOCUS       sequence-227295-       17333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent voltage sensor ASAP2s cassette into the mouse
            TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17333)
  AUTHORS   Daigle TL, Madisen L, Hage TA, Valley MT, Knoblich U, Larsen RS, 
            Takeno MM, Huang L, Gu H, Larsen R, Mills M, Bosma-Moody A, Siverts 
            LA, Walker M, Graybuck LT, Yao Z, Fong O, Nguyen TN, Garren E, Lenz 
            GH, Chavarha M, Pendergraft J, Harrington J, Hirokawa KE, Harris JA,
            Nicovich PR, McGraw MJ, Ollerenshaw DR, Smith KA, Baker CA, Ting JT,
            Sunkin SM, Lecoq J, Lin MZ, Boyden ES, Murphy GJ, da Costa NM, 
            Waters J, Li L, Tasic B, Zeng H
  TITLE     A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced 
            Brain-Cell-Type Targeting and Functionality.
  JOURNAL   Cell. 2018 Jul 12;174(2):465-480.e22. doi: 
            10.1016/j.cell.2018.06.035.
  PUBMED    30007418
REFERENCE   2  (bases 1 to 17333)
  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.cell.2018.06"; journalName: "Cell"; date: "2018-07-12- 
            12"; volume: "174"; issue: "2"; pages: "465-480.e22"
COMMENT     Sequence Label: Ai170 (TIT2L-ASAP2s-Kv-ICL-tTA2) targeting vector
FEATURES             Location/Qualifiers
     source          1..17333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    414..447
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     polyA_signal    2975..3056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3000..3019
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(3070..3088,3089..3111,3112..3130,3131..3153,3154..3172,
                     3173..3195,3196..3214,3215..3237,3238..3256,3257..3279,
                     3280..3298,3299..3321,3322..3340)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3373..3411
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3408..3432
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3545..3578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3655..3703
                     /note="synthetic polyadenylation signal"
     polyA_signal    3712..4188
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    4200..4661
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(4688..4721)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6271..6859
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6742..6753)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6890..7097
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        9621..9924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9926..10203
                     /label=chicken beta-actin promoter
     intron          10204..11220
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     11144..11166
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     11228..11247
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    11308..11341
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     polyA_signal    11379..11427
                     /note="synthetic polyadenylation signal"
     polyA_signal    11436..11912
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11947..11966)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11991..12039
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    12218..12251
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             join(12276..12893,12894..12899,12900..12935,12936..12938,
                     12939..12974,12975..12977,12978..13013,13014..13022)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    13065..13653
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(13536..13547)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    13684..13891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    13905..13938
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        13965..14464
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(14000..14023)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     14397..14416
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(15112..15128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15112..15128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15125..15147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15136..15152
                     /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(15160..15166,15167..15184,15185..15190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15205..15226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15343..15360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15514..16102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15594..15613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16273..17064,17065..17133))
                     /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     16896..16915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17134..17238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17306..17324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accgaagttc
      421 ctattcttca aatagtatag gaacttcttt cggccggcca atcgattgga tccgtttaat
      481 tcgagctcgc tagagcccca tcctcactga ctccgtcctg gagttggatg agagataatg
      541 gccttacgtt gtgccagggg agggtcgggc tggatttagc aagatttacc ttctccaaag
      601 agcggtgctg cagtggcaca gctgcccacg gaggtggggg ggtcaccgtc cctggaggtg
      661 atgaagaact gtggggatgt ggcactgagg gacatggcca gtgggcacgg tgggtgggtt
      721 ggggttggtc ttggggatct tggagggctt ttccagcctt catgatttga cgattgtatg
      781 aacatctaca tggcaattct ccagctgcct gtcccagtcc tactgaccca gctgtatctc
      841 tccaggcaag ctcttccacc ccttctgctt gcatccagac accatcaaac atgcaggctc
      901 agacacaggg accagcagtg tctgtggcct ttttgtgctc ctctccatgc tgggttttaa
      961 cttgctcttt gtccttctat cctatcttct tatccttaag gctgttctga acgctgtgac
     1021 ttggagagtg tcccagagcc ctcaacacct gcatgtccca cgtccatgct gtcctgcact
     1081 tccttatccc caagatctgc ctctccgtga tgcactgaat tggcaaacat gtgtcacccc
     1141 agaccaacaa tgtcacagca aactccccct tgataggaca agggggaatg gctttacact
     1201 gagacagggg aggtttgggt tggatatgag gaggcagttt ttcccccaga gggtggtgac
     1261 gcactgaaca ggttgcccaa ggaggctgtg gatgccccat ccctgcaggc attcaaggcc
     1321 aggctggatg tggctctggg cagcctgggc tgctggttga tgaccctgca catagcaggg
     1381 ggttggatct ggatgagcac tgtgctcctt tgcaacccag gccgttctat gattctgtca
     1441 ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc
     1501 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     1561 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     1621 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     1681 ttgggggggg gctgtccctc tagcgagctc ggtacccaga gccccatcct cactgactcc
     1741 gtcctggagt tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga
     1801 tttagcaaga tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg
     1861 tgggggggtc accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca
     1921 tggccagtgg gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc
     1981 agccttcatg atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc
     2041 cagtcctact gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat
     2101 ccagacacca tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt
     2161 gtgctcctct ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc
     2221 cttaaggctg ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat
     2281 gtcccacgtc catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca
     2341 ctgaattggc aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat
     2401 aggacaaggg ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg
     2461 cagtttttcc cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg
     2521 ccccatccct gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct
     2581 ggttgatgac cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca
     2641 acccaggccg ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc
     2701 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg
     2761 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg
     2821 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga
     2881 gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctagg ggtacccggg
     2941 gatccaccgg tatcgattcg gccggccgac gtcggcagtg aaaaaaatgc tttatttgtg
     3001 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagtttcga
     3061 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     3121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     3181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     3241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     3301 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     3361 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     3421 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     3481 gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc aaattcaatt
     3541 aaatataact tcgtatagca tacattatac gaagttattc gcgatgaata aatgaaagct
     3601 tgcagatccc ctcagcacca tggctagcgg cagcctcgga gtttgaatag atagaataaa
     3661 atatctttat tttcattcca tctgtgtgtt ggttttttgt gtgagatcta cgggtggcat
     3721 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3781 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3841 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3901 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3961 atctccgcct cctgggttca agcgattctc ctgcctcagg ctcccgagtt gttgggattc
     4021 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4081 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4141 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gcctcagcgc
     4201 tgtaagtctg cagaaattga tgatctatta aacaataaag atgtccacta aaatggaagt
     4261 ttttcctgtc atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag
     4321 gattggagct acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa
     4381 ggctctttac tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc
     4441 aaaaccaaat taagggccag ctcattcctc ccactcatga tctatagatc tatagatctc
     4501 tcgtgggatc attgtttttc tcttgattcc cactttgtgg ttctaagtac tgtggtttcc
     4561 aaatgtgtca gtttcatagc ctgaagaacg agatcagcag cctctgttcc acatacactt
     4621 cattctcagt attgttttgc caagttctaa ttccatcaga aagcttttaa ttaactgaat
     4681 agctaacata acttcgtata gcatacatta tacgaagtta tcacgcgtat ggagacgact
     4741 gtgaggtatg aacaggggtc agagctcact aaaacttcga gctctccaac agcagatgag
     4801 cccacgataa agattgatga tggtcgtgat gagggtaatg aacaagacag ctgttccaat
     4861 accattagga gaaaaatttc cccgtttgtg atgtcatttg gattcagagt atttggagtt
     4921 gtgcttatca ttgtagacat catagtggtg attgtggatc tggccatcag tgagaagaaa
     4981 agaggcatta gagagattct tgaaggtgtt tccctggcta tagcactctt cttccttgtt
     5041 gatgttctca tgagagtgtt tgttgaaggc ttcaagaact atttccggtc caaactgaat
     5101 actttggatg cagtcatagt agtgggcact ctgctaatta atatgaccta ctccttctct
     5161 gaccttgctg cctttaacag ccataacgtg tatattaccg cggataaaca gaaaaacggc
     5221 attaaagcga actttaccgt gcgccataac gtggaagatg gcagcgtgca gctggcggat
     5281 cattatcagc agaacacccc gattggcgat ggcccggtgc tgctgccgga taaccattat
     5341 ctgagcaccc agaccgtgct gagcaaagat ccgaacgaaa aacgcgatca catggtgctg
     5401 ctggaatttg tgaccgcagc gggcattaca cacggcatgg atgaactgta tggcggcacc
     5461 ggcggcagcg cgagccaggg cgaagaactg tttaccggcg tggtgccgat tctggtggaa
     5521 ctggatggcg atgtgaacgg ccataaattt agcgtgcgcg gcgaaggcga aggcgatgcg
     5581 accattggca aactgaccct gaaatttatt tgcaccaccg gcaaactacc ggtgccgtgg
     5641 ccgaccctgg tgaccacctt aacctatggc gtgcagtgct ttagccgcta tccggatcat
     5701 atgaaacgcc atgatttttt taaaagcgcg atgccggaag gctatgtgca ggaacgcacc
     5761 attagcttta aagatgatgg caaatataaa acccgcgcgg tggtgaaatt tgaaggcgat
     5821 accctggtga accgcattga actgaaaggc accgatttta aagaagatgg caacattctg
     5881 gggcataaac tggaatataa cacagatcag atgccgcaga tggttactct tcttcgagtt
     5941 ctgagaattg ttatcttaat aagaatattt cagctggctt cacagaagaa acaacttgaa
     6001 gtggtaaccc ttaagggttc ttcaggttca tctggatctt ccacgcgtca gagccagccg
     6061 attctgaaca ccaaagaaat ggcgccgcag agcaaaccgc cggaagaact ggaaatgagc
     6121 agcatgccga gcccggtggc gccgctgccg gcgcgcaccg aaggcgtgat tgatatgcgc
     6181 agcatgagca gcattgatag ctttattagc tgcgcgaccg attttccgga agcgacccgc
     6241 ttttgaacgc gttacgtaaa gcttgtcgat aatcaacctc tggattacaa aatttgtgaa
     6301 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6361 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6421 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6481 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6541 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6601 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6661 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     6721 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6781 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6841 ctttgggccg cctccccgca tcgacaccga gcgctgctcg aggcctcgac tgtgccttct
     6901 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6961 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     7021 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     7081 agcaggcatg ctggggacac gtgggatcct ctagagcccc atcctcactg actccgtcct
     7141 ggagttggat gagagataat ggccttacgt tgtgccaggg gagggtcggg ctggatttag
     7201 caagatttac cttctccaaa gagcggtgct gcagtggcac agctgcccac ggaggtgggg
     7261 gggtcaccgt ccctggaggt gatgaagaac tgtggggatg tggcactgag ggacatggcc
     7321 agtgggcacg gtgggtgggt tggggttggt cttggggatc ttggagggct tttccagcct
     7381 tcatgatttg acgattgtat gaacatctac atggcaattc tccagctgcc tgtcccagtc
     7441 ctactgaccc agctgtatct ctccaggcaa gctcttccac cccttctgct tgcatccaga
     7501 caccatcaaa catgcaggct cagacacagg gaccagcagt gtctgtggcc tttttgtgct
     7561 cctctccatg ctgggtttta acttgctctt tgtccttcta tcctatcttc ttatccttaa
     7621 ggctgttctg aacgctgtga cttggagagt gtcccagagc cctcaacacc tgcatgtccc
     7681 acgtccatgc tgtcctgcac ttccttatcc ccaagatctg cctctccgtg atgcactgaa
     7741 ttggcaaaca tgtgtcaccc cagaccaaca atgtcacagc aaactccccc ttgataggac
     7801 aagggggaat ggctttacac tgagacaggg gaggtttggg ttggatatga ggaggcagtt
     7861 tttcccccag agggtggtga cgcactgaac aggttgccca aggaggctgt ggatgcccca
     7921 tccctgcagg cattcaaggc caggctggat gtggctctgg gcagcctggg ctgctggttg
     7981 atgaccctgc acatagcagg gggttggatc tggatgagca ctgtgctcct ttgcaaccca
     8041 ggccgttcta tgattctgtc attctaaatc tctctttcag cctaaagctt tttccccgta
     8101 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc
     8161 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg
     8221 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac
     8281 gtaattacat ccctgggggc tttggggggg ggctgtccct ctagagcccc atcctcactg
     8341 actccgtcct ggagttggat gagagataat ggccttacgt tgtgccaggg gagggtcggg
     8401 ctggatttag caagatttac cttctccaaa gagcggtgct gcagtggcac agctgcccac
     8461 ggaggtgggg gggtcaccgt ccctggaggt gatgaagaac tgtggggatg tggcactgag
     8521 ggacatggcc agtgggcacg gtgggtgggt tggggttggt cttggggatc ttggagggct
     8581 tttccagcct tcatgatttg acgattgtat gaacatctac atggcaattc tccagctgcc
     8641 tgtcccagtc ctactgaccc agctgtatct ctccaggcaa gctcttccac cccttctgct
     8701 tgcatccaga caccatcaaa catgcaggct cagacacagg gaccagcagt gtctgtggcc
     8761 tttttgtgct cctctccatg ctgggtttta acttgctctt tgtccttcta tcctatcttc
     8821 ttatccttaa ggctgttctg aacgctgtga cttggagagt gtcccagagc cctcaacacc
     8881 tgcatgtccc acgtccatgc tgtcctgcac ttccttatcc ccaagatctg cctctccgtg
     8941 atgcactgaa ttggcaaaca tgtgtcaccc cagaccaaca atgtcacagc aaactccccc
     9001 ttgataggac aagggggaat ggctttacac tgagacaggg gaggtttggg ttggatatga
     9061 ggaggcagtt tttcccccag agggtggtga cgcactgaac aggttgccca aggaggctgt
     9121 ggatgcccca tccctgcagg cattcaaggc caggctggat gtggctctgg gcagcctggg
     9181 ctgctggttg atgaccctgc acatagcagg gggttggatc tggatgagca ctgtgctcct
     9241 ttgcaaccca ggccgttcta tgattctgtc attctaaatc tctctttcag cctaaagctt
     9301 tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga
     9361 aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg
     9421 atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg
     9481 ggggagggac gtaattacat ccctgggggc tttggggggg ggctgtccct ctagagtcga
     9541 cggcgcgccc tgcaggaatc tagttattaa tagtaatcaa ttacggggtc attagttcat
     9601 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     9661 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     9721 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     9781 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     9841 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     9901 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     9961 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
    10021 gcgatggggg cggggggggg gggggggcgc gcgccrggsg gggsggggsg gggsgrgggg
    10081 sggggsgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
    10141 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
    10201 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
    10261 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
    10321 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
    10381 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
    10441 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
    10501 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
    10561 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
    10621 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
    10681 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
    10741 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
    10801 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
    10861 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
    10921 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
    10981 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
    11041 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
    11101 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
    11161 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
    11221 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgatttga
    11281 taccgcgggc cccgcgtgga cggtaccata acttcgtata aagtatccta tacgaagtta
    11341 tccatggcta gcggcagcct cggagtttga atagatagaa taaaatatct ttattttcat
    11401 tccatctgtg tgttggtttt ttgtgtgaga tctacgggtg gcatccctgt gacccctccc
    11461 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
    11521 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
    11581 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
    11641 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
    11701 ttcaagcgat tctcctgcct caggctcccg agttgttggg attccaggca tgcatgacca
    11761 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
    11821 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
    11881 aggcgtgaac cactgctccc ttccctgtcc ttctgcctca gctaattgag taggggggag
    11941 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
    12001 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
    12061 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
    12121 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
    12181 tcggggcggc aggccgtgga agttcctatt ctctagaata acttcgtata aagtatccta
    12241 tacgaagtta tacgcgcgcg tccatttaat tcaccatgtc tagactggac aagagcaaag
    12301 tcataaactc tgctctggaa ttactcaatg aagtcggtat cgaaggcctg acgacaagga
    12361 aactcgctca aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc
    12421 gggccctgct cgatgccctg gcaatcgaga tgctggacag gcatcatacc cacttctgcc
    12481 ccctggaagg cgagtcatgg caagactttc tgcggaacaa cgccaagtca ttccgctgtg
    12541 ctctcctctc acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac
    12601 agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga
    12661 acgcactgta cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggatc
    12721 aggagcatca agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac
    12781 ttctgagaca agcaattgag ctgttcgacc atcagggagc cgaacctgcc ttccttttcg
    12841 gcctggaact aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccgg
    12901 ccgacgccct tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg
    12961 accttgatat gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt
    13021 aactaagtaa ggatcaaatg gcgcgttacg taaagcttgt cgataatcaa cctctggatt
    13081 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    13141 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    13201 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    13261 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    13321 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    13381 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    13441 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
    13501 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    13561 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    13621 cgagtcggat ctccctttgg gccgcctccc cgcatcgaca ccgagcgctg ctcgaggcct
    13681 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
    13741 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
    13801 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
    13861 attgggaaga caatagcagg catgctgggg acacgtggga tccggtgcca gggcgtgccc
    13921 ttgggctccc cgggcgcgtt tgggcggccg catcgaattc taccgggtag gggaggcgct
    13981 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
    14041 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
    14101 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
    14161 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
    14221 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
    14281 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
    14341 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
    14401 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
    14461 ttcgacctgc aatcgccgct agccgccacc atgaaaaagc ctgaactcac cgcgacgtct
    14521 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
    14581 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
    14641 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
    14701 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
    14761 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
    14821 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
    14881 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
    14941 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
    15001 agtgtaagtg ctagccccta ggcctgaatc gaagttccta ttcttcaaaa ggtataggaa
    15061 cttcgagctc gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct
    15121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    15181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    15241 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    15301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    15361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    15421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    15481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    15541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    15601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    15661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    15721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    15781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    15841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    15901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    15961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    16021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    16081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    16141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    16201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    16261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    16321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    16381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    16441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    16501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    16561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    16621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    16681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    16741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    16801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    16861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    16921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    16981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    17041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    17101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    17161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    17221 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
    17281 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
