LOCUS       sequence-222568-        8816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible suppression of KRAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8816)
  AUTHORS   Shao DD, Xue W, Krall EB, Bhutkar A, Piccioni F, Wang X, Schinzel 
            AC, Sood S, Rosenbluh J, Kim JW, Zwang Y, Roberts TM, Root DE, Jacks
            T, Hahn WC
  TITLE     KRAS and YAP1 Converge to Regulate EMT and Tumor Survival.
  JOURNAL   Cell. 2014 Jul 3;158(1):171-84. doi: 10.1016/j.cell.2014.06.004. 
            Epub 2014 Jun 19.
  PUBMED    24954536
REFERENCE   2  (bases 1 to 8816)
  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.2014.06.004"; journalName: "Cell"; date: "2014-07-3-
            3"; volume: "158"; issue: "1"; pages: "171-84"
COMMENT     Sequence Label: Tet-pLKO-puro shKRAS
FEATURES             Location/Qualifiers
     source          1..8816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        275..369
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     286..305
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    323..341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    351..369
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    470..587
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        636..1146
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1065..1083
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     intron          1155..1727
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1735..1754
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        1782..1800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1811..2431
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1884..1903)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    2466..3039
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2633..2650)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2860..2879
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3059..3658
                     /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(3059..3078)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3555..3575
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3786..4019
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4091..4212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4128..4147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4182..4201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4252..4387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4408..4426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4436..4453)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4436..4452)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4445..4467)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4594..5049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4681..4700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4891..4912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5075..5179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5180..5248,5249..6040)
                     /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(5398..5417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6211..6799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6700..6719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6953..6970
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7087..7108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7123..7128,7129..7146,7147..7153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7161..7177
                     /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     7166..7188
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7185..7201
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7185..7201
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7220..7240
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7222..7240
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7268..7494
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7495..7675
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7722..7847
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8340..8573
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8758..8802
                     /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"
ORIGIN
        1 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
       61 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
      121 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
      181 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
      241 gtatcgatca cgagactagc ctcgagcggc cgcaatattt gcatgtcgct atgtgttctg
      301 ggaaatcacc ataaacgtga aatccctatc agtgatagag acttataagt tccctatcag
      361 tgatagagac accggcagtt gagaccttct aattggctcg agccaattag aaggtctcaa
      421 ctgtttttga attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa
      481 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
      541 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
      601 gggacagcag agatccactt tggccgcggc tcgagggggt tggggttgcg ccttttccaa
      661 ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc
      721 ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc
      781 gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt
      841 tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc
      901 ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc
      961 aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag
     1021 gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc
     1081 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
     1141 ccccaggggg atctgtgagt ttggggaccc ttgattgttc tttctttttc gctattgtaa
     1201 aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc
     1261 ccttgtatca ccatggaccc tcatgataat tttgtttctt tcactttcta ctctgttgac
     1321 aaccattgtc tcctcttatt ttcttttcat tttctgtaac tttttcgtta aactttagct
     1381 tgcatttgta acgaattttt aaattcactt ttgtttattt gtcagattgt aagtactttc
     1441 tctaatcact tttttttcaa ggcaatcagg gtatattata ttgtacttca gcacagtttt
     1501 agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata aattctggct
     1561 ggcgtggaaa tattcttatt ggtagaaaca actacatcct ggtcatcatc ctgcctttct
     1621 ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct gagtccaaac
     1681 cgggcccctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1741 tgctggttat tgtgctgtct catcattttg gcaaagaatt gtaatacgac tcactatagg
     1801 gcgaattgat atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct
     1861 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga
     1921 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat
     1981 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga
     2041 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc
     2101 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt
     2161 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt
     2221 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga
     2281 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt
     2341 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt
     2401 agaaaaacaa cttaaatgtg aaagtgggtc cgcgtacagc ggctcccggg agttctaggg
     2461 atctgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     2521 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     2581 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     2641 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     2701 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     2761 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     2821 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2881 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2941 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     3001 cacggggacg tggttttcct ttgaaaaaca cgatgataag gatccaccgg agcttaccat
     3061 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3121 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3181 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3241 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3301 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3361 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3421 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     3481 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     3541 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     3601 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     3661 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc
     3721 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     3781 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     3841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     3901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4021 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     4081 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4141 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4201 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     4441 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4501 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     4561 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4801 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4861 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4921 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4981 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5041 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5101 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5161 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5221 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5281 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5341 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5401 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5461 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5521 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5581 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5641 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5701 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5761 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5821 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5881 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5941 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6001 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6841 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6901 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6961 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7021 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7081 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     7141 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     7201 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     7261 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     7321 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     7381 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     7441 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     7501 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7561 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7621 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     7681 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     7741 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     7801 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     7861 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     7921 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     7981 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     8041 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     8101 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     8161 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     8221 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     8281 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     8341 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     8401 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     8461 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     8521 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     8581 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     8641 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     8701 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     8761 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaa
//
