LOCUS       sequence-232859-       12222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For targeted integration and inducible expression of human NODAL 
            using doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12222)
  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 12222)
  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-NODAL
FEATURES             Location/Qualifiers
     source          1..12222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        84..462
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    92..110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    128..146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    164..182
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    200..218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    236..254
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    272..290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    308..326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     371..395
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1678..1743
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1873..1893
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2318..2439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2355..2374)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2409..2428
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2477..2701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2690..2707
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2739..3338)
                     /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(2822..2842)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3319..3338
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3348..3401)
                     /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    3425..3450
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    3457..4260
                     /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(4299..4317)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4338..4354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4362..4378
                     /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(4386..4392,4393..4410,4411..4416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4431..4452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4569..4586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4740..5328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4820..4839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5452..5520,5521..6312)
                     /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(5670..5689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(6673..6692)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7287..7306
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(7709..8011)
                     /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(7811..7830)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     8134..8156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8148..8165
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8149..8165
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8172..8191
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8172..8190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8225..9061
                     /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    9135..9156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9171..9176,9177..9194,9195..9201)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9209..9225
                     /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     9233..9249
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9592..9611
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    9610..9665
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     9711..9730
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(9821..10567)
                     /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"
     primer_bind     complement(10605..10624)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          10632..11640
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(10686..10708)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(11641..11916)
                     /label=chicken beta-actin promoter
     enhancer        11919..12222
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
       61 gttttcgtct tcaagaattc ctcgagttta ctccctatca gtgatagaga acgtatgaag
      121 agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga
      181 gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct
      241 atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc
      301 agtttactcc ctatcagtga tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct
      361 ataaaagcag agctcgttta gtgaaccgtc agatcgcctg gagcaattcc acaacacttt
      421 tgtcttatac caactttccg taccacttcc taccctcgta aagtcgaccc accatgcacg
      481 cccactgcct gcccttcctt ctgcacgcct ggtgggccct actccaggcg ggtgctgcga
      541 cggtggccac tgcgctcctg cgtacgcggg ggcagccctc gtcgccatcc cctctggcgt
      601 acatgctgag cctctaccgc gacccgctgc cgagggcaga catcatccgc agcctacagg
      661 cagaagatgt ggcagtggat gggcagaact ggacgtttgc ttttgacttc tccttcctga
      721 gccaacaaga ggatctggca tgggctgagc tccggctgca gctgtccagc cctgtggacc
      781 tccccactga gggctcactt gccattgaga ttttccacca gccaaagccc gacacagagc
      841 aggcttcaga cagctgctta gagcggtttc agatggacct attcactgtc actttgtccc
      901 aggtcacctt ttccttgggc agcatggttt tggaggtgac caggcctctc tccaagtggc
      961 tgaagcaccc tggggccctg gagaagcaga tgtccagggt agctggagag tgctggccgc
     1021 ggccccccac accgcctgcc accaatgtgc tccttatgct ctactccaac ctctcgcagg
     1081 agcagaggca gctgggtggg tccaccttgc tgtgggaagc cgagagctcc tggcgggccc
     1141 aggagggaca gctgtcctgg gagtggggca agaggcaccg gcgacatcac ttgccagaca
     1201 gaagtcaact gtgtcggaag gtcaagttcc aggtggactt caacctgatc ggatggggct
     1261 cctggatcat ctaccccaag cagtacaacg cctatcgctg tgagggcgag tgtcctaatc
     1321 ctgttgggga ggagtttcat ccgaccaacc atgcatacat ccagagtctg ctgaaacgtt
     1381 accagcccca ccgagtccct tccacttgtt gtgccccagt gaagaccaag ccgctgagca
     1441 tgctgtatgt ggataatggc agagtgctcc tagatcacca taaagacatg atcgtggaag
     1501 aatgtgggtg cctctgaacg cgtcatatgg ctagcctgca gggatccaat gtaactgtat
     1561 tcagcgatga cgaaattctt agctattgta atactctaga ggatctttgt gaaggaacct
     1621 tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta
     1681 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt
     1741 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     1801 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     1861 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     1921 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     1981 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     2041 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
     2101 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
     2161 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat
     2221 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2281 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2341 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2401 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgcg gcctagtcaa
     2461 taatcaatgt cgagacccat agagcccacc gcatccccag catgcctgct attgtcttcc
     2521 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     2581 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     2641 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     2701 gtcgaggctg atcagcgggt ttaaacgggc cctctagatc aggcaccggg cttgcgggtc
     2761 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     2821 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     2881 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     2941 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     3001 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     3061 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     3121 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     3181 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     3241 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     3301 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatct cgagcctagg gccgggattc
     3361 tcctccacgt caccgcatgt tagaagactt cctctgccct ctccgctgcc agatctctcg
     3421 aggccctgtg ggaggaagag aagaggtcag aagcttgccc cactgtgggg tggaggggac
     3481 agataaaagt acccagaacc agagccacat taaccggccc tgggaatata aggtggtccc
     3541 agctcgggga cacaggatcc ctggaggcag caaacatgct gtcctgaagt ggacataggg
     3601 gcccgggttg gaggaagaag actagctgag ctctcggacc cctggaagat gccatgacag
     3661 ggggctggaa gagctagcac agactagaga ggtaaggggg gtaggggagc tgcccaaatg
     3721 aaaggagtga gaggtgaccc gaatccacag gagaacgggg tgtccaggca aagaaagcaa
     3781 gaggatggag aggtggctaa agccagggag acggggtact ttggggttgt ccagaaaaac
     3841 ggtgatgatg caggcctaca agaaggggag gcgggacgca agggagacat ccgtcggaga
     3901 aggccatcct aagaaacgag agatggcaca ggccccagaa ggagaaggaa aagggaaccc
     3961 agcgagtgaa gacggcatgg ggttgggtga gggaggagag atgcccggag aggacccaga
     4021 cacggggagg atccgctcag aggacatcac gtggtgcagc gccgagaagg aagtgctccg
     4081 gaaagagcat ccttgggcag caacacagca gagagcaagg ggaagaggga gtggaggaag
     4141 acggaacctg aaggaggcgg cagggaagga tctgggccag ccgtagaggt gacccaggcc
     4201 acaagctgca gacagaaagc ggcacaggcc caggggagag aatgctggtc agagaaagca
     4261 aagggcgaat tcgtttaaac ctgcaggact agctagtccc tttagtgagg gttaattctg
     4321 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5401 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     5761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6001 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6061 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6301 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6361 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gagcggatac
     6421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6481 gtgccacctg atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6541 gaaattgtaa gcgttaataa ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     6601 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     6661 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     6721 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     6781 gcaggcatcg ccatgcatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     6841 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     6901 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     6961 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     7021 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     7081 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     7141 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     7201 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     7261 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     7321 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     7381 tctcttgatc agagcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     7441 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     7501 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     7561 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     7621 tccggggtca gcaccgtttc tgcggactgg ctttctacgt gaaaaggatc taggtgaaga
     7681 tcctttttga taatctcatg cctgacattt atattcccca gaacatcagg ttaatggcgt
     7741 ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg
     7801 gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg
     7861 ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg gggatcacca
     7921 tccgtcgccc cggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc
     7981 tctctctttt ataggtgtaa accttaaact gccgtacgta taggctgcgc aactgttggg
     8041 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     8101 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     8161 ccagtgaatt gtaatacgac tcactatagg gcgaattgaa tttagcggcc gcgaattcgc
     8221 ccttagagca gagccaggaa cccctgtagg gaaggggcag gagagccagg ggcatgagat
     8281 ggtggacgag gaagggggac agggaagcct gagcgcctct cctgggcttg ccaaggactc
     8341 aaacccagaa gcccagagca gggccttagg gaagcgggac cctgctctgg gcggaggaat
     8401 atgtcccaga tagcactggg gactctttaa ggaaagaagg atggagaaag agaaagggag
     8461 tagaggcggc cacgacctgg tgaacaccta ggacgcacca ttctcacaaa gggagttttc
     8521 cacacggaca cccccctcct caccacagcc ctgccaggac ggggctggct actggcctta
     8581 tctcacaggt aaaactgacg cacggaggaa caatataaat tggggactag aaaggtgaag
     8641 agccaaagtt agaactcagg accaacttat tctgattttg tttttccaaa ctgcttctcc
     8701 tcttgggaag tgtaaggaag ctgcagcacc aggatcagtg aaacgcacca gacagccgcg
     8761 tcagagcagc tcaggttctg ggagagggta gcgcagggtg gccactgaga accgggcagg
     8821 tcacgcatcc cccccttccc tcccaccccc tgccaagctc tccctcccag gatcctctct
     8881 ggctccatcg taagcaaacc ttagaggttc tggcaaggag agagatggct ccaggaaatg
     8941 ggggtgtgtc accagataag gaatctgcct aacaggaggt gggggttaga cccaatatca
     9001 ggagactagg aaggaggagg cctaaggatg gggcttttct gtcaccaatc ctgtccctag
     9061 taaagcttag tactgtcgat ccgctggcac gacaggtttc ccgactggaa agcgggcagt
     9121 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     9181 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     9241 agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc ataagagaag
     9301 agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta gtaaaacatg
     9361 taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat
     9421 ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg acagcagggg gctgtttcat
     9481 atactgatga cctctttata gccaaccttt gttcatggca gccagcatat gggcatatgt
     9541 tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc tcccatatgt
     9601 ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa taaatttcct
     9661 ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat ttttggcaga
     9721 gggaaaaaga tctcagtggt atttgtgagc cagggcattg gccacaccag ccaccacctt
     9781 ctgataggca gcctgcacct gaggagtgaa ttcacgcgcc ttacccgggg agcatgtcaa
     9841 ggtcaaaatc gtcaagagcg tcagcaggca gcatatcaag gtcaaagtcg tcaagggcat
     9901 cggctgggag catgtctaag tcaaaatcgt caagggcgtc ggtcggcccg ccgctttcgc
     9961 actttagctg tttctccagg ccacatatga ttagttccag gccgaaaagg aaggcaggtt
    10021 cggctccctg ccggtcgaac agctcaattg cttgtttcag aagtgggggc atagaatcgg
    10081 tggtaggtgt ctctctttcc tcttttgcta cttgatgctc ctgttcctcc aatacgcagc
    10141 ccagtgtaaa gtggcccacg gcggacagag cgtacagtgc gttctccagg gagaagcctt
    10201 gctgacacag gaacgcgagc tgattttcca gggtttcgta ctgtttctct gttgggcggg
    10261 tgccgagatg cactttagcc ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg
    10321 cgttgttccg cagaaagtct tgccatgact cgccttccag ggggcaggag tgggtatgat
    10381 gcctgtccag catctcgatt ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc
    10441 agtacagggt aggctgctca actcccagct tttgagcgag tttccttgtc gtcaggcctt
    10501 cgataccgac tccattgagt aattccagag cagagtttat gactttgctc ttgtccagtc
    10561 tagacatggt gctcgacgaa ttctttgcca aaatgatgag acagcacaat aaccagcacg
    10621 ttgcccagga gctgtaggaa aaagaagaag gcatgaacat ggttagcaga ggctctagag
    10681 ccgccggtca cacgccagaa gccgaacccc gccctgcccc gtcccccccg aaggcagccg
    10741 tccccctgcg gcagccccga ggctggagat ggagaagggg acggcggcgc ggcgacgcac
    10801 gaaggccctc cccgcccatt tccttcctgc cggcgccgca ccgcttcgcc cgcgcccgct
    10861 agagggggtg cggcggcgcc tcccagattt cggctccgcc agatttggga caaaggaagt
    10921 ccctgcgccc tctcgcacga ttaccataaa aggcaatggc tgcggctcgc cgcgcctcga
    10981 cagccgccgg cgctccgggg ccgccgcgcc cctcccccga gccctccccg gcccgaggcg
    11041 gccccgcccc gcccggcacc cccacctgcc gccacccccc gcccggcacg gcgagccccg
    11101 cgccacgccc cgcacggagc cccgcacccg aagccgggcc gtgctcagca actcggggag
    11161 gggggtgcag gggggggtta cagcccgacc gccgcgccca caccccctgc tcaccccccc
    11221 acgcacacac cccgcacgca gcctttgttc ccctcgcagc ccccccgcac cgcggggcac
    11281 cgcccccggc cgcgctcccc tcgcgcacac gcggagcgca caaagccccg cgccgcgccc
    11341 gcagcgctca cagccgccgg gcagcgcggg ccgcacgcgg cgctccccac gcacacacac
    11401 acgcacgcac cccccgagcc gctccccccc gcacaaaggg ccctcccgga gccctttaag
    11461 gctttcacgc agccacagaa aagaaacgag ccgtcattaa accaagcgct aattacagcc
    11521 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
    11581 gggcgggcgg cgcgaggcgg cgcggagcgg ggcacggggc gaaggcaacg cagcgactcc
    11641 cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg ccataaaagg
    11701 aaactttcgg agcgcgccgc tctgattggc tgccgccgca cctctccgcc tcgccccgcc
    11761 ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc cccccccccc gcccccatcg
    11821 ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag gggggggaga
    11881 tggggagagt gaagcagaac gtggggctca cctcgaccca tggtaatagc gatgactaat
    11941 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
    12001 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
    12061 tgtactgcca agtgggcagt ttaccgtaaa tagtccaccc attgacgtca atggaaagtc
    12121 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
    12181 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cg
//
