Tet-pLKO-puro

LOCUS       Tet-pLKO-puro      10633 bp    DNA     circular SYN 14-APR-2025
DEFINITION  3rd generation lentiviral plasmid for inducible expression of shRNA;
            puromycin selection. See manual for detailed protocols. No further 
            technical support is available from the depositing laboratory..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10633)
  AUTHORS   Wiederschain D, Wee S, Chen L, Loo A, Yang G, Huang A, Chen Y, 
            Caponigro G, Yao YM, Lengauer C, Sellers WR, Benson JD
  TITLE     Single-vector inducible lentiviral RNAi system for oncology target 
            validation.
  JOURNAL   Cell Cycle. 2009 Feb 1. 8(3):498-504.
  PUBMED    19177017
REFERENCE   2  (bases 1 to 10633)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Cycle.
            2009 Feb 1. 8(3):498-504."
COMMENT     Sequence Label: Tet-pLKO-puro
FEATURES             Location/Qualifiers
     source          1..10633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..59
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    176..197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(212..217,218..235,236..242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    250..266
                     /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     255..277
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     274..290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     274..290
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     309..329
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        311..329
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        357..583
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             584..764
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    811..936
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1429..1662
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1847..1891
                     /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"
     promoter        2180..2274
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     2191..2210
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    2228..2246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2256..2274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    4192..4309
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4358..4868
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4787..4805
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     intron          4877..5449
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     5457..5476
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        5504..5522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5533..6153
                     /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(5606..5625)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    6188..6761
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6355..6372)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6582..6601
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6781..7380
                     /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(6781..6800)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7277..7297
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7508..7741
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7813..7934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7850..7869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7904..7923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7974..8109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(8130..8148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8158..8175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8158..8174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8167..8189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8316..8771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8403..8422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8613..8634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8797..8901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8902..8970,8971..9762)
                     /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(9120..9139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9933..10521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10422..10441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
       61 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      181 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      241 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      301 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
      361 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
      421 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
      481 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
      541 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
      601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
      961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1261 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     1321 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1381 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     1441 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     1501 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     1561 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     1621 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     1681 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1741 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     1801 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     1861 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     1921 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     1981 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     2041 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     2101 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga
     2161 ctagcctcga gcggccgcaa tatttgcatg tcgctatgtg ttctgggaaa tcaccataaa
     2221 cgtgaaatcc ctatcagtga tagagactta taagttccct atcagtgata gagacaccgg
     2281 tgtgttgtaa atgagcacac aaaatacaca tgctaaaata ttatattcta tgacctttat
     2341 aaaatcaacc aaaatcttct ttttaataac tttagtatca ataattagaa tttttatgtt
     2401 cctttttgca aacttttaat aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc
     2461 gcgttgtttt cttcaccttt aataatagct actccaccac ttgttcctaa gcggtcagct
     2521 cctgcttcaa tcattttttg agcatcttca aatgttctaa ctccaccagc tgctttaact
     2581 aaagcattgt ctttaacaac tgacttcatt agtttaacat cttcaaatgt tgcacctgat
     2641 tttgaaaatc ctgttgatgt tttaacaaat tctaatccag cttcaacagc tatttcacaa
     2701 gctttcatga tttcttcttt tgttaataaa caattttcca taatacattt aacaacatgt
     2761 gatccagctg ctttttttac agctttcatg tcttctaaaa ctaattcata atttttgtct
     2821 tttaatgcac caatatttaa taccatatca atttctgttg caccatcttt aattgcttca
     2881 gaaacttcga atgcttttgt agctgttgtg catgcaccta gaggaaaacc tacaacattt
     2941 gttattccta catttgtgcc ttttaataat tctttacaat agcttgttca atatgaatta
     3001 acacaaactg ttgcaaaatc aaattcaatt gcttcatcac ataattgttt aatttcagct
     3061 ttcgtagcat cttgttttaa taatgtgtga tctatatatt tgtttagttt cattttttct
     3121 cctatatatt catttttaat tttaattctt taataatttc gtctacttta actttagcgt
     3181 tttgaacaga ttcaccaaca cctataaaat aaatttttag tttaggttca gttccacttg
     3241 ggcgaacagc aaatcatgac ttatcttcta aataaaattt tagtaagtct tgtcctggca
     3301 tattatacat tccatcgatg tagtcttcaa cattaacaac tttaagtcca gcaatttgag
     3361 ttaagggtgt tgctctcaat gatttcatta atggttcaat ttttaatttc ttttcttctg
     3421 gtttaaaatt caagtttaaa gtgaaagtgt aatatgcacc catttcttta aataaatctt
     3481 ctaaatagtc tactaatgtt ttattttgtt ttttataaaa tcaagcagcc tctgctatta
     3541 atatagaagc ttgtattcca tctttatctc tagctgagtc atcaattaca tatccataac
     3601 tttcttcata agcaaaaaca aaatttaatc cgttatcttc ttctttagca atttctctac
     3661 ccattcattt aaatccagtt aaagttttta caatattaac tccatatttt tcatgagcga
     3721 ttctatcacc caaatcactt gttacaaaac ttgaatatag agccggattt tttggaatgc
     3781 tatttaagcg ttttagattt gataattttc aatcaattaa aattggtcct gtttgatttc
     3841 catctaatct tacaaaatga ccatcatgtt ttattgccat tccaaatctg tcagcatctg
     3901 ggtcattcat aataataata tctgcatcat gtttaatacc atattcaagc ggtatttttc
     3961 atgcaggatc aaattctgga tttggattta caacattttt aaatgtttca tcttcaaatg
     4021 catgctcttc aacctcaata acgttatatc ctgattcacg taatattttt ggggtaaatt
     4081 tagttcctgt tccattaact gcgctaaaaa taatttttaa atctttttta gcttcttgct
     4141 cttttttgta gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
     4201 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     4261 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     4321 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
     4381 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     4441 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     4501 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     4561 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     4621 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     4681 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     4741 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     4801 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     4861 ctccccaggg ggatctgtga gtttggggac ccttgattgt tctttctttt tcgctattgt
     4921 aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg
     4981 tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc tactctgttg
     5041 acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt taaactttag
     5101 cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt gtaagtactt
     5161 tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt cagcacagtt
     5221 ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata taaattctgg
     5281 ctggcgtgga aatattctta ttggtagaaa caactacatc ctggtcatca tcctgccttt
     5341 ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact ctgagtccaa
     5401 accgggcccc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     5461 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgtaatacg actcactata
     5521 gggcgaattg atatgtctag attagataaa agtaaagtga ttaacagcgc attagagctg
     5581 cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta
     5641 gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc
     5701 attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa
     5761 gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga
     5821 gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa
     5881 ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct
     5941 gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa
     6001 gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta
     6061 tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga
     6121 ttagaaaaac aacttaaatg tgaaagtggg tccgcgtaca gcggctcccg ggagttctag
     6181 ggatctgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     6241 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     6301 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     6361 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     6421 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     6481 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     6541 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     6601 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     6661 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     6721 accacgggga cgtggttttc ctttgaaaaa cacgatgata aggatccacc ggagcttacc
     6781 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     6841 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     6901 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     6961 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     7021 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7081 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7141 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     7201 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7261 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7321 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7381 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta
     7441 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     7501 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     7561 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7621 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7681 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7741 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7801 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7921 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     7981 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     8041 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     8101 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     8161 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8221 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8281 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     8341 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     8401 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     8461 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     8521 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     8581 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     8641 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     8701 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     8761 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8881 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     8941 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     9001 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     9061 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     9121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     9181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     9241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     9481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9601 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     9661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     9781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     9841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     9901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    10021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    10081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    10141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    10201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    10261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    10321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    10381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    10441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    10501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    10621 attaccgcct ttg
//