LOCUS       sequence-232861-       12195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For targeted integration and inducible expression of a human NODAL 
            splice variant using doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12195)
  AUTHORS   Findlay SD, Bilyk O, Lypka K, Waskiewicz AJ, Postovit L
  TITLE     Genetically regulated human NODAL splice variants are differentially
            post-transcriptionally processed and functionally distinct
  JOURNAL   (2018) bioRxiv 276170
REFERENCE   2  (bases 1 to 12195)
  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: "(2018) 
            bioRxiv 276170"
COMMENT     Sequence Label: AAVS1-TRE3G-NODALvar
FEATURES             Location/Qualifiers
     source          1..12195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..409
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    39..57
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    75..93
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    111..129
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    147..165
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    183..201
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    219..237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    255..273
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     318..342
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1598..1663
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1793..1813
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2238..2359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2275..2294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2329..2348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2397..2621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2610..2627
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2659..3258)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2742..2762)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3239..3258
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3268..3321)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3345..3370
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    3377..4180
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(4219..4237)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4258..4274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4282..4298
                     /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(4306..4312,4313..4330,4331..4336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4489..4506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4660..5248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4740..4759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5372..5440,5441..6232)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5590..5609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(6593..6612)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7207..7226
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(7629..7931)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(7731..7750)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     8054..8076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8068..8085
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8069..8085
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8092..8111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8092..8110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8145..8981
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    9055..9076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9091..9096,9097..9114,9115..9121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9129..9145
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9153..9169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9512..9531
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    9530..9585
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     9631..9650
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(9741..10487)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     intron          10552..11560
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(10606..10628)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(11561..11836)
                     /label=chicken beta-actin promoter
     enhancer        11839..12142
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ataactagtt ttcgtcttca agaattcctc gagtttactc cctatcagtg atagagaacg
       61 tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac tccctatcag
      121 tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta tgaccagttt
      181 actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg atagagaacg
      241 tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc gtgtacggtg
      301 ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag caattccaca
      361 acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag tcgacccacc
      421 atgcacgccc actgcctgcc cttccttctg cacgcctggt gggccctact ccaggcgggt
      481 gctgcgacgg tggccactgc gctcctgcgt acgcgggggc agccctcgtc gccatcccct
      541 ctggcgtaca tgctgagcct ctaccgcgac ccgctgccga gggcagacat catccgcagc
      601 ctacaggcag aagatgtggc agtggatggg cagaactgga cgtttgcttt tgacttctcc
      661 ttcctgagcc aacaagagga tctggcatgg gctgagctcc ggctgcagct gtccagccct
      721 gtggacctcc ccactgaggg ctcacttgcc attgagattt tccaccagcc aaagcccgac
      781 acagagcagg cttcagacag ctgcttagag cggtttcaga tggacctatt cactgtcact
      841 ttgtcccagg tcaccttttc cttgggcagc atggttttgg aggtgaccag gcctctctcc
      901 aagtggctga agcaccctgg ggccctggag aagcagatgt ccagggtagc tggagagtgc
      961 tggccgcggc cccccacacc gcctgccacc aatgtgctcc ttatgctcta ctccaacctc
     1021 tcgcaggagc agaggcagct gggtgggtcc accttgctgt gggaagccga gagctcctgg
     1081 cgggcccagg agggacagct gtcctgggag tggggcaaga ggcaccggcg acatcacttg
     1141 ccagacagaa gtcaactgtg tcggaaggtc aagttccagg tggacttcaa cctgatcgga
     1201 tggggctcct ggatcatcta ccccaagcag tacaacgcct atcgctgtga gggcgagtgt
     1261 cctaatcctg ttggggagga gtttcatccg accaaccatg catacatcca ggtggccctg
     1321 ccctgctgtc caaggtcata tgggaccaaa atgttttcat tttactccat gaagtctgga
     1381 atgagaattt cttggacttg caatatatcc tccatgccaa gcctcagagt ctgctgaacg
     1441 cgtcatatgg ctagcctgca gggatccaat gtaactgtat tcagcgatga cgaaattctt
     1501 agctattgta atactctaga ggatctttgt gaaggaacct tacttctgtg gtgtgacata
     1561 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     1621 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     1681 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
     1741 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
     1801 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
     1861 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
     1921 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
     1981 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
     2041 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
     2101 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
     2161 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2221 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2341 aactcatcaa tgtatcttat catgtctgcg gcctagtcaa taatcaatgt cgagacccat
     2401 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2461 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2521 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2581 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt
     2641 ttaaacgggc cctctagatc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc
     2701 ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt
     2761 gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc
     2821 ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt
     2881 ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg
     2941 ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac
     3001 cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc
     3061 gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt
     3121 gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa
     3181 cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt
     3241 gggcttgtac tcggtcatct cgagcctagg gccgggattc tcctccacgt caccgcatgt
     3301 tagaagactt cctctgccct ctccgctgcc agatctctcg aggccctgtg ggaggaagag
     3361 aagaggtcag aagcttgccc cactgtgggg tggaggggac agataaaagt acccagaacc
     3421 agagccacat taaccggccc tgggaatata aggtggtccc agctcgggga cacaggatcc
     3481 ctggaggcag caaacatgct gtcctgaagt ggacataggg gcccgggttg gaggaagaag
     3541 actagctgag ctctcggacc cctggaagat gccatgacag ggggctggaa gagctagcac
     3601 agactagaga ggtaaggggg gtaggggagc tgcccaaatg aaaggagtga gaggtgaccc
     3661 gaatccacag gagaacgggg tgtccaggca aagaaagcaa gaggatggag aggtggctaa
     3721 agccagggag acggggtact ttggggttgt ccagaaaaac ggtgatgatg caggcctaca
     3781 agaaggggag gcgggacgca agggagacat ccgtcggaga aggccatcct aagaaacgag
     3841 agatggcaca ggccccagaa ggagaaggaa aagggaaccc agcgagtgaa gacggcatgg
     3901 ggttgggtga gggaggagag atgcccggag aggacccaga cacggggagg atccgctcag
     3961 aggacatcac gtggtgcagc gccgagaagg aagtgctccg gaaagagcat ccttgggcag
     4021 caacacagca gagagcaagg ggaagaggga gtggaggaag acggaacctg aaggaggcgg
     4081 cagggaagga tctgggccag ccgtagaggt gacccaggcc acaagctgca gacagaaagc
     4141 ggcacaggcc caggggagag aatgctggtc agagaaagca aagggcgaat tcgtttaaac
     4201 ctgcaggact agctagtccc tttagtgagg gttaattctg agcttggcgt aatcatggtc
     4261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5101 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gagcggatac atatttgaat gtatttagaa
     6361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg
     6421 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaataa
     6481 ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     6541 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     6601 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     6661 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgcatgg
     6721 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     6781 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     6841 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     6901 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     6961 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     7021 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     7081 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     7141 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     7201 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     7261 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agagcttgat
     7321 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     7381 ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc
     7441 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     7501 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     7561 tgcggactgg ctttctacgt gaaaaggatc taggtgaaga tcctttttga taatctcatg
     7621 cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     7681 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     7741 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     7801 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc cggcgtgtca
     7861 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     7921 accttaaact gccgtacgta taggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7981 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8041 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     8101 tcactatagg gcgaattgaa tttagcggcc gcgaattcgc ccttagagca gagccaggaa
     8161 cccctgtagg gaaggggcag gagagccagg ggcatgagat ggtggacgag gaagggggac
     8221 agggaagcct gagcgcctct cctgggcttg ccaaggactc aaacccagaa gcccagagca
     8281 gggccttagg gaagcgggac cctgctctgg gcggaggaat atgtcccaga tagcactggg
     8341 gactctttaa ggaaagaagg atggagaaag agaaagggag tagaggcggc cacgacctgg
     8401 tgaacaccta ggacgcacca ttctcacaaa gggagttttc cacacggaca cccccctcct
     8461 caccacagcc ctgccaggac ggggctggct actggcctta tctcacaggt aaaactgacg
     8521 cacggaggaa caatataaat tggggactag aaaggtgaag agccaaagtt agaactcagg
     8581 accaacttat tctgattttg tttttccaaa ctgcttctcc tcttgggaag tgtaaggaag
     8641 ctgcagcacc aggatcagtg aaacgcacca gacagccgcg tcagagcagc tcaggttctg
     8701 ggagagggta gcgcagggtg gccactgaga accgggcagg tcacgcatcc cccccttccc
     8761 tcccaccccc tgccaagctc tccctcccag gatcctctct ggctccatcg taagcaaacc
     8821 ttagaggttc tggcaaggag agagatggct ccaggaaatg ggggtgtgtc accagataag
     8881 gaatctgcct aacaggaggt gggggttaga cccaatatca ggagactagg aaggaggagg
     8941 cctaaggatg gggcttttct gtcaccaatc ctgtccctag taaagcttag tactgtcgat
     9001 ccgctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     9061 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     9121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     9181 caagcttggg ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga
     9241 gtagtcagga gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg
     9301 ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt
     9361 ttctatggaa taaggaatgg acagcagggg gctgtttcat atactgatga cctctttata
     9421 gccaaccttt gttcatggca gccagcatat gggcatatgt tgccaaactc taaaccaaat
     9481 actcattctg atgttttaaa tgatttgccc tcccatatgt ccttccgagt gagagacaca
     9541 aaaaattcca acacactatt gcaatgaaaa taaatttcct ttattagcca gaagtcagat
     9601 gctcaagggg cttcatgatg tccccataat ttttggcaga gggaaaaaga tctcagtggt
     9661 atttgtgagc cagggcattg gccacaccag ccaccacctt ctgataggca gcctgcacct
     9721 gaggagtgaa ttcacgcgcc ttacccgggg agcatgtcaa ggtcaaaatc gtcaagagcg
     9781 tcagcaggca gcatatcaag gtcaaagtcg tcaagggcat cggctgggag catgtctaag
     9841 tcaaaatcgt caagggcgtc ggtcggcccg ccgctttcgc actttagctg tttctccagg
     9901 ccacatatga ttagttccag gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac
     9961 agctcaattg cttgtttcag aagtgggggc atagaatcgg tggtaggtgt ctctctttcc
    10021 tcttttgcta cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg
    10081 gcggacagag cgtacagtgc gttctccagg gagaagcctt gctgacacag gaacgcgagc
    10141 tgattttcca gggtttcgta ctgtttctct gttgggcggg tgccgagatg cactttagcc
    10201 ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg cgttgttccg cagaaagtct
    10261 tgccatgact cgccttccag ggggcaggag tgggtatgat gcctgtccag catctcgatt
    10321 ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt aggctgctca
    10381 actcccagct tttgagcgag tttccttgtc gtcaggcctt cgataccgac tccattgagt
    10441 aattccagag cagagtttat gactttgctc ttgtccagtc tagacatggt gctcgacgaa
    10501 ttctttgcca aaatgatgag acagcacaac aaccagcacg ttgcccagga gctgtaggaa
    10561 aaagaagaag gcatgaacat ggttagcaga ggctctagag ccgccggtca cacgccagaa
    10621 gccgaacccc gccctgcccc gtcccccccg aaggcagccg tccccctgcg gcagccccga
    10681 ggctggagat ggagaagggg acggcggcgc ggcgacgcac gaaggccctc cccgcccatt
    10741 tccttcctgc cggcgccgca ccgcttcgcc cgcgcccgct agagggggtg cggcggcgcc
    10801 tcccagattt cggctccgcc agatttggga caaaggaagt ccctgcgccc tctcgcacga
    10861 ttaccataaa aggcaatggc tgcggctcgc cgcgcctcga cagccgccgg cgctccgggg
    10921 ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc gcccggcacc
    10981 cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc cgcacggagc
    11041 cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag gggggggtta
    11101 cagcccgacc gccgcgccca caccccctgc tcaccccccc acgcacacac cccgcacgca
    11161 gcctttgttc ccctcgcagc ccccccgcac cgcggggcac cgcccccggc cgcgctcccc
    11221 tcgcgcacac gcggagcgca caaagccccg cgccgcgccc gcagcgctca cagccgccgg
    11281 gcagcgcggg ccgcacgcgg cgctccccac gcacacacac acgcacgcac cccccgagcc
    11341 gctccccccc gcacaaaggg ccctcccgga gccctttaag gctttcacgc agccacagaa
    11401 aagaaacgag ccgtcattaa accaagcgct aattacagcc cggaggagaa gggccgtccc
    11461 gcccgctcac ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg
    11521 cgcggagcgg ggcacggggc gaaggcaacg cagcgactcc cgcccgccgc gcgcttcgct
    11581 ttttataggg ccgccgccgc cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc
    11641 tctgattggc tgccgccgca cctctccgcc tcgccccgcc ccgcccctcg ccccgccccg
    11701 ccccgcctgg cgcgcgcccc cccccccccc gcccccatcg ctgcacaaaa taattaaaaa
    11761 ataaataaat acaaaattgg gggtggggag gggggggaga tggggagagt gaagcagaac
    11821 gtggggctca cctcgaccca tggtaatagc gatgactaat acgtagatgt actgccaagt
    11881 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
    11941 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
    12001 ttaccgtaaa tagtccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
    12061 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
    12121 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
    12181 aattgattac tatta
//
