LOCUS       sequence-402530-       13456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-ON 3rd generation expression vector with inducible expression of
            BCL9 and a puromycin selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13456)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 13456)
  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: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: pLVX-TetOne-Puro-BCL9
FEATURES             Location/Qualifiers
     source          1..13456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..376
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    4859..4993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5037..5152
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(5415..5459)
                     /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    5644..5877
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6374..6499
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6546..7179
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7347..7481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        7529..7633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7634..7702,7703..8494)
                     /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(7852..7871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8665..9253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9154..9173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9407..9424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9541..9562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9577..9582,9583..9600,9601..9607)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9615..9631
                     /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     9620..9642
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9639..9655
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9639..9655
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             9783..10416
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10623..11211
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             10729..10740
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     11138..11158
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11225..11824)
                     /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(11308..11328)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     11805..11824
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(11833..12162)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11847..11982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(12173..12919)
                     /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"
     promoter        complement(12938..13448)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13001..13019)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      361 ttcctaccct cgtaaagaat tcggcatgca ttccagtaac cctaaagtga ggagctctcc
      421 atcaggaaac acacagagta gccctaagtc aaagcaggag gtgatggtcc gtccccctac
      481 agtgatgtcc ccatctggaa acccccagct ggattccaaa ttctccaatc agggtaaaca
      541 ggggggctca gccagccaat cccagccatc cccctgtgac tccaagagtg ggggccatac
      601 ccctaaagca ctccctggcc caggtgggag catggggctg aagaatgggg ctggaaatgg
      661 tgccaagggc aaggggaaaa gggagcgaag tatttccgcc gactcctttg atcagagaga
      721 tcctgggact ccaaacgatg actctgacat taaagaatgt aattctgctg accacataaa
      781 gtcccaggat tcccagcaca caccacactc gatgacccca tcaaatgcta cagcccccag
      841 gtcttctacc ccctcccatg gccaaactac tgccacagag cccacacctg ctcagaagac
      901 tccagccaaa gtggtgtacg tgttttctac tgagatggcc aataaagctg cagaagctgt
      961 tttgaagggc caggttgaaa ctatcgtctc tttccacatc cagaacattt ctaacaacaa
     1021 gacagagaga agcacagcgc ctctgaacac acagatatct gcccttcgga atgatccgaa
     1081 acctctccca caacagcccc cagctccggc caaccaggac cagaattctt cccagaatac
     1141 cagactgcag ccaactccac ccattccggc accagcaccc aagcctgccg cacccccacg
     1201 tcccctggac cgggagagtc ctggggtaga aaacaaactg attccttctg taggaagtcc
     1261 tgccagctcc actccactgc ccccagatgg tactgggccc aactcaactc ccaacaatag
     1321 ggcagtgacc cctgtctccc aggggagcaa tagctcttca gcagatccca aagcccctcc
     1381 gcctccacca gtgtccagtg gcgagccccc cacactggga gagaatcccg atggcctatc
     1441 tcaggagcag ctggagcacc gggagcgctc cttacaaact ctcagagata tccagcgcat
     1501 gctttttcct gatgagaaag aattcacagg agcacaaagt gggggaccgc agcagaatcc
     1561 tggggtatta gatgggcctc agaaaaaacc agaagggcca atacaggcca tgatggccca
     1621 atcccaaagc ctaggtaagg gacctgggcc ccggacagac gtgggagctc catttggccc
     1681 tcaaggacat agagatgtac ccttttctcc agatgaaatg gttccacctt ctatgaactc
     1741 ccagtctggg accataggac ccgaccacct tgaccatatg actcccgagc agatagcgtg
     1801 gctgaaactg cagcaggagt tttatgaaga gaagaggagg aagcaggaac aagtggttgt
     1861 ccagcagtgt tccctccagg acatgatggt ccatcagcac gggcctcggg gagtggtccg
     1921 aggacccccc cctccatacc agatgacccc tagtgaaggc tgggcacctg ggggtacaga
     1981 gccattttct gatggtatca acatgccaca ttctctgccc ccgaggggca tggctcccca
     2041 ccccaacatg ccagggagcc agatgcgcct ccctggattt gcaggcatga taaactctga
     2101 aatggaaggg ccgaatgtcc ccaaccctgc atctagacca ggtctttctg gagtcagttg
     2161 gccagatgat gtgccaaaaa tcccagatgg tcgaaatttt cctcctggcc agggcatttt
     2221 cagcggtcct ggccgagggg aacgcttccc aaacccccaa ggattgtctg aagagatgtt
     2281 tcagcagcag ctggcagaga aacagctggg tctcccccca gggatggcca tggaaggcat
     2341 caggcccagc atggagatga acaggatgat tccaggctcc cagcgccaca tggagcctgg
     2401 gaataacccc attttccctc gaataccagt tgagggccct ctgagtcctt ctaggggtga
     2461 ctttccaaaa ggaattcccc cacagatggg ccctggtcgg gaacttgagt ttgggatggt
     2521 tcctagtggg atgaagggag atgtcaatct aaatgtcaac atgggatcca actctcagat
     2581 gatacctcag aagatgagag aggctggggc gggccctgag gagatgctga aattacgccc
     2641 aggtggctca gacatgctgc ctgctcagca gaagatggtg ccactgccat ttggtgagca
     2701 cccccagcag gagtatggca tgggccccag accattcctt cccatgtctc agggtccagg
     2761 cagcaacagt ggcttgcgga atctcagaga accaattggg cccgaccaga ggactaacag
     2821 ccggctcagt catatgccac cactacctct caacccttcc agtaacccca ccagcctcaa
     2881 cacagctcct ccagttcagc gcggcctggg gcggaagccc ttggatatat ctgtggcagg
     2941 cagccaggtg cattccccag gcattaaccc tctgaagtct cccacgatgc accaagtcca
     3001 gtcaccaatg ctgggctcgc cctcggggaa cctcaagtcc ccccagactc catcgcagct
     3061 ggcaggcatg ctggcgggcc cagctgctgc tgcttccatt aagtcccccc ctgttttggg
     3121 gtctgctgct gcttcacctg tccacctcaa gtctccatca cttcctgccc cgtcacctgg
     3181 atggacctct tctccaaaac ctccccttca gagtcctggg atccctccaa accataaagc
     3241 acccctcacc atggcctccc cagccatgct gggaaatgta gagtcaggtg gccccccacc
     3301 tcctacagcc agccagcctg cctctgtgaa tatccctgga agtcttccct ctagtacacc
     3361 ttataccatg cctccagagc caaccctttc ccagaaccca ctctctatta tgatgtctcg
     3421 aatgtccaag tttgcaatgc ccagttccac cccgttatac catgatgcta tcaagactgt
     3481 ggccagctca gatgacgact cccctccagc tcgttctccc aacttgccat caatgaataa
     3541 tatgccagga atgggcatta atacacagaa tcctcgaatt tcaggtccaa accccgtggt
     3601 tccgatgcca accctcagcc caatgggaat gacccagcca ctttctcact ccaatcagat
     3661 gccctctcca aatgccgtgg gacccaacat acctcctcat ggggtcccaa tggggcctgg
     3721 cttgatgtca cacaatccta tcatggggca tgggtcccag gagccaccga tggtacctca
     3781 aggacggatg ggcttccccc agggcttccc tccagtacag tctcccccac agcaggttcc
     3841 attccctcac aatggcccca gtggggggca gggcagcttc ccaggaggga tgggtttccc
     3901 aggagaaggc ccccttggcc gccccagcaa cctgccccaa agttcagcag atgcagcact
     3961 ttgcaagcct ggaggccccg ggggtcctga ctccttcact gtcctgggga acagcatgcc
     4021 ttcggtgttt acagacccag atctgcagga ggtcatccga cctggagcca ccggaatacc
     4081 tgagtttgat ctatcccgca ttattccatc tgagaagccc agccagacgc tgcaatattt
     4141 ccctcgaggg gaagttccag gccgtaaaca gccccagggt cctggacctg ggttttcaca
     4201 catgcagggg atgatgggcg aacaagcccc cagaatggga ctagcattac ctggcatggg
     4261 aggtccaggg ccagtgggaa ctccggacat ccctcttggt acagctccat ccatgccagg
     4321 ccacaacccc atgagaccac cagcctttct ccaacaaggc atgatgggac ctcaccatcg
     4381 gatgatgtca ccagcacaat ctacaatgcc cggccagccc accctgatga gcaatccagc
     4441 tgctgccgtg ggcatgattc ctggcaagga tcgggggcct gccgggctct acacccaccc
     4501 tgggcctgtg ggctctccag gcatgatgat gtccatgcag ggcatgatgg gaccccaaca
     4561 gaacatcatg atccccccac agatgaggcc ccggggcatg gctgctgacg tgggcatggg
     4621 tggatttagc caaggacctg gcaacccagg aaacatgatg ttttaaaccg gtgccggcgg
     4681 atccagacca cctcccctgc gagctaagct ggacagccaa tgacgggtaa gagagtgaca
     4741 tttttcacta acctaagaca ggagggccgt cagagctact gcctaatcca aagacgggta
     4801 aaagtgataa aaatgtatca ctccaaccta agacaggcgc agcttccgag ggatttgaga
     4861 tccagacatg ataagataca ttgatgagtt tggacaaacc aaaactagaa tgcagtgaaa
     4921 aaaatgcctt atttgtgaaa tttgtgatgc tattgcctta tttgtaacca ttataagctg
     4981 caataaacaa gttaagtcga taaactggat ctctgctgtc cctgtaataa acccgaaaat
     5041 tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt
     5101 ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta aaggcgatac
     5161 cgtcgagatc cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct
     5221 tctattcctt cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat
     5281 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     5341 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     5401 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     5461 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     5521 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     5581 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     5641 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     5701 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     5761 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     5821 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     5881 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     5941 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     6001 tctcctcctc caggtctgaa gatcagcggc cggccgcttg ctgtgcggtg gtcttacttt
     6061 tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt
     6121 gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg
     6181 tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca
     6241 ggccaggatt aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt
     6301 tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc
     6361 cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa
     6421 tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc
     6481 gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc
     6541 gccactgcta gagattttcc acactgacta aaagggtctg agggatctct agttaccaga
     6601 gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt
     6661 aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag
     6721 agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc
     6781 cagtcccgcc caggccacgc ctccctggaa agtccccagc ggaaagtccc ttgtagcaag
     6841 ctcgatatca gcagttcttg aagtactccg gatgcagctc tcgggccacg tgatgaaatg
     6901 ctaggcggct gtcaaacctc cactctaaca cttctctctc cgggtcatcc atcccatgca
     6961 ggctcacagg gtgtaacaag ctggtgttct ctcctttatt ggcctcttct accttatctg
     7021 gctcaactgg tactagcttg tagcaccatc caaaggtcag tggatatctg acccctggcc
     7081 ctggtgtgta gttctgctaa tcagggaagt agccttgtgt gtggtagatc cacagatcaa
     7141 ggatatcttg tcttctttgg gagtgaatta gcccttccaa ctactaagtt tgtagtacat
     7201 atttaacaaa tacaatttct ttaaaatgaa aataattcag aggaatcaca ggtttagagt
     7261 aaatgaaacc acaggtaatt ggcagtggta atagggtatg gggtgggaag tttgggatga
     7321 ttttggttag cttgagttat ccagttgatc cagacatgat aagatacatt gatgagtttg
     7381 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     7441 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tgatctcccg atccgtcgac
     7501 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7561 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7621 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7681 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7741 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7801 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7861 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7921 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7981 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     8041 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     8101 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8161 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8221 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8281 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8341 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8401 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8461 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8521 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9361 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9421 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     9481 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     9541 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     9601 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     9661 ttacgccaag ctctagctag aggtcgacgg taaacgctag caatgtcaag gcctctcact
     9721 ctctgatatt catttctttg caagttataa atactgaata ataagatgac atgaactact
     9781 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     9841 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     9901 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     9961 cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc cactccccag
    10021 tcccgcccag gccacgcctc cctggaaagt ccccagcgga aagtcccttg tagcaagctc
    10081 gatatcagca gttcttgaag tactccggat gcagctctcg ggccacgtga tgaaatgcta
    10141 ggcggctgtc aaacctccac tctaacactt ctctctccgg gtcatccatc ccatgcaggc
    10201 tcacagggtg taacaagctg gtgttctctc ctttattggc ctcttctacc ttatctggct
    10261 caactggtac tagcttgtag caccatccaa aggtcagtgg atatctgacc cctggccctg
    10321 gtgtgtagtt ctgctaatca gggaagtagc cttgtgtgtg gtagatccac agatcaagga
    10381 tatcttgtct tcgttgggag tgaattagcc cttccagtcc cctcttttct tttaaaaagt
    10441 ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa aggtacctga ggtgtgactg
    10501 gaaaacccac ctcctcctcc tcttgtgctt ctagccaggc acaatcagca ttggtagctg
    10561 ctgtattgct acttgtgatt gctccatgtt tttctaggtc tcgactgcag aattaattcc
    10621 aggcggggag gcggcccaaa gggagatccg actcgtctga gggcgaaggc gaagacgcgg
    10681 aagaggccgc agagccggca gcaggccgcg ggaaggaagg tccgctggat tgagggccga
    10741 agggacgtag cagaaggacg tcccgcgcag aatccaggtg gcaacacagg cgagcagcca
    10801 tggaaaggac gtcagcttcc ccgacaacac cacggaattg tcagtgccca acagccgagc
    10861 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa tagggagggg
    10921 gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca atgccccaac cagtgggggt
    10981 tgcgtcagca aacacagtgc acaccacgcc acgttgcctg acaacgggcc acaactcctc
    11041 ataaagagac agcaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc
    11101 aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata
    11161 gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tgttccagac
    11221 gcgttcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
    11281 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
    11341 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
    11401 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
    11461 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
    11521 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
    11581 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
    11641 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
    11701 ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
    11761 tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
    11821 tcatggtcgc gttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
    11881 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
    11941 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
    12001 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
    12061 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
    12121 tgctggggag cctggggact ttccacaccc taactgacac acattcgcgc gtttacccgg
    12181 ggagcatgtc aaggtcaaaa tcgtcaagag cgtcagcagg cagcatatca aggtcaaagt
    12241 cgtcaagggc atcggctggg agcatgtcta agtcaaaatc gtcaagggcg tcggtcggcc
    12301 cgccgctttc gcactttagc tgtttctcca ggccacatat gattagttcc aggccgaaaa
    12361 ggaaggcagg ttcggctccc tgccggtcga acagctcaat tgcttgtttc agaagtgggg
    12421 gcatagaatc ggtggtaggt gtctctcttt cctcttttgc tacttgatgc tcctgttcct
    12481 ccaatacgca gcccagtgta aagtggccca cggcggacag agcgtacagt gcgttctcca
    12541 gggagaagcc ttgctgacac aggaacgcga gctgattttc cagggtttcg tactgtttct
    12601 ctgttgggcg ggtgccgaga tgcactttag ccccgtcgcg atgtgagagg agagcacagc
    12661 ggtatgactt ggcgttgttc cgcagaaagt cttgccatga ctcgccttcc agggggcagg
    12721 agtgggtatg atgcctgtcc agcatctcga ttggcagggc atcgagcagg gcccgcttgt
    12781 tcttcacgtg ccagtacagg gtaggctgct caactcccag cttttgagcg agtttccttg
    12841 tcgtcaggcc ttcgataccg actccattga gtaattccag agcagagttt atgactttgc
    12901 tcttgtccag tctagacatg gtaattcgat gatccccctg gggagagagg tcggtgattc
    12961 ggtcaacgag ggagccgact gccgacgtgc gctccggagg cttgcagaat gcggaacacc
    13021 gcgcgggcag gaacagggcc cacactaccg ccccacaccc cgcctcccgc accgcccctt
    13081 cccggccgct gctctcggcg cgccctgctg agcagccgct attggccaca gcccatcgcg
    13141 gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc gagggtacta gtgagacgtg
    13201 cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa ggaaccttcc cgacttaggg
    13261 gcggagcagg aagcgtcgcc ggggggccca caagggtagc ggcgaagatc cgggtgacgc
    13321 tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc gccgctgcgt ttcccggaac
    13381 cacgcccaga gcagccgcgt ccctgcgcaa acccagggct gccttggaaa aggcgcaacc
    13441 ccaaccccgt ggaatt
//
