LOCUS       sequence-204300-       10223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive Flag-tagged SMAD4 expression in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10223)
  AUTHORS   David CJ, Huang YH, Chen M, Su J, Zou Y, Bardeesy N, 
            Iacobuzio-Donahue CA, Massague J
  TITLE     TGF-beta Tumor Suppression through a Lethal EMT.
  JOURNAL   Cell. 2016 Feb 25;164(5):1015-30. doi: 10.1016/j.cell.2016.01.009. 
            Epub 2016 Feb 18.
  PUBMED    26898331
REFERENCE   2  (bases 1 to 10223)
  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.2016.01.009"; journalName: "Cell"; date: 
            "2016-02-25- 25"; volume: "164"; issue: "5"; pages: "1015-30"
COMMENT     Sequence Label: pLVX-IRES Hyg SMAD4
FEATURES             Location/Qualifiers
     source          1..10223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2201..2504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2659..2679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2705..2729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2818..2827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2827..2850
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(4547..5121,5122..5124,5125..5133)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4714..4731)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4941..4960
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5159..6178
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             5159..6178
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    6192..6780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6245..6265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6663..6674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6987..7620
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7748..7764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7748..7764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7761..7783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7772..7788
                     /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(7796..7802,7803..7820,7821..7826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7841..7862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7979..7996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8150..8738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8230..8249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8909..9700,9701..9769))
                     /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     9532..9551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9770..9874)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9922..10056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9959..9978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10013..10032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcct cggatccgcc
     2821 accatggatt acaaggacga cgatgacaag atggacaata tgtctattac gaatacacca
     2881 acaagtaatg atgcctgtct gagcattgtg catagtttga tgtgccatag acaaggtgga
     2941 gagagtgaaa catttgcaaa aagagcaatt gaaagtttgg taaagaagct gaaggagaaa
     3001 aaagatgaat tggattcttt aataacagct ataactacaa atggagctca tcctagtaaa
     3061 tgtgttacca tacagagaac attggatggg aggcttcagg tggctggtcg gaaaggattt
     3121 cctcatgtga tctatgcccg tctctggagg tggcctgatc ttcacaaaaa tgaactaaaa
     3181 catgttaaat attgtcagta tgcgtttgac ttaaaatgtg atagtgtctg tgtgaatcca
     3241 tatcactacg aacgagttgt atcacctgga attgatctct caggattaac actgcagagt
     3301 aatgctccat caagtatgat ggtgaaggat gaatatgtgc atgactttga gggacagcca
     3361 tcgttgtcca ctgaaggaca ttcaattcaa accatccagc atccaccaag taatcgtgca
     3421 tcgacagaga catacagcac cccagctctg ttagccccat ctgagtctaa tgctaccagc
     3481 actgccaact ttcccaacat tcctgtggct tccacaagtc agcctgccag tatactgggg
     3541 ggcagccata gtgaaggact gttgcagata gcatcagggc ctcagccagg acagcagcag
     3601 aatggattta ctggtcagcc agctacttac catcataaca gcactaccac ctggactgga
     3661 agtaggactg caccatacac acctaatttg cctcaccacc aaaacggcca tcttcagcac
     3721 cacccgccta tgccgcccca tcccggacat tactggcctg ttcacaatga gcttgcattc
     3781 cagcctccca tttccaatca tcctgctcct gagtattggt gttccattgc ttactttgaa
     3841 atggatgttc aggtaggaga gacatttaag gttccttcaa gctgccctat tgttactgtt
     3901 gatggatacg tggacccttc tggaggagat cgcttttgtt tgggtcaact ctccaatgtc
     3961 cacaggacag aagccattga gagagcaagg ttgcacatag gcaaaggtgt gcagttggaa
     4021 tgtaaaggtg aaggtgatgt ttgggtcagg tgccttagtg accacgcggt ctttgtacag
     4081 agttactact tagacagaga agctgggcgt gcacctggag atgctgttca taagatctac
     4141 ccaagtgcat atataaaggt ctttgatttg cgtcagtgtc atcgacagat gcagcagcag
     4201 gcggctactg cacaagctgc agcagctgcc caggcagcag ccgtggcagg aaacatccct
     4261 ggcccaggat cagtaggtgg aatagctcca gctatcagtc tgtcagctgc tgctggaatt
     4321 ggtgttgatg accttcgtcg cttatgcata ctcaggatga gttttgtgaa aggctgggga
     4381 ccggattacc caagacagag catcaaagaa acaccttgct ggattgaaat tcacttacac
     4441 cgggccctcc agctcctaga cgaagtactt cataccatgc cgattgcaga cccacaacct
     4501 ttagactgag aattcgagac tagttctaga gcggccgcgg atcccgcccc tctccctccc
     4561 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     4621 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4681 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     4741 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     4801 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4861 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4921 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     4981 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5041 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5101 tttgaaaaac acgatgataa gcttgccaca acccgtacca aagatggata gatccggaaa
     5161 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     5221 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     5281 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     5341 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     5401 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     5461 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     5521 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     5581 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     5641 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5701 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     5761 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5821 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5881 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     5941 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     6001 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     6061 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     6121 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagac
     6181 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6241 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6301 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6361 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6421 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6481 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6541 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     6601 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6661 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6721 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6781 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     6841 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     6901 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6961 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga
     7021 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     7081 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     7141 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     7201 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     7261 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     7321 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     7381 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     7441 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7501 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7561 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7621 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7681 agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag agcttggcgt
     7741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     8041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     8101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     8161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     9001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     9061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     9121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     9181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     9241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     9301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     9361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9901 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
     9961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10021 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    10081 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    10141 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    10201 atatgtacta caaacttagt agt
//
