LOCUS       Exported                8849 bp ds-DNA     circular SYN 14-FEB-2022
DEFINITION  Tet/Dox inducible shRNA lentivirus with blasticidin selection.
ACCESSION   .
VERSION     .
KEYWORDS    EZ-Tet-pLKO-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8849)
  AUTHORS   Frank SB, Schulz VV, Miranti CK
  TITLE     A streamlined method for the design and cloning of shRNAs into an 
            optimized Dox-inducible lentiviral vector.
  JOURNAL   BMC Biotechnol. 2017 Feb 28;17(1):24. doi: 
            10.1186/s12896-017-0341-x.
  PUBMED    28245848
REFERENCE   2  (bases 1 to 8849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 14, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    35..608
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(202..219)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     429..448
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             626..1024
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             1219..1452
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1524..1645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1561..1580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1615..1634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1685..1820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1747..1766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        complement(1841..1859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1869..1886)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1869..1885)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1878..1900)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2027..2482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2114..2133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2324..2345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2508..2612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2613..3473
                     /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(2831..2850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3644..4232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4133..4152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4386..4403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4520..4541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4556..4586
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4594..4610
                     /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     4599..4621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4618..4634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4618..4634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4653..4673
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4655..4673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4701..4927
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4928..5108
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5155..5280
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5773..6006
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6191..6235
                     /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        6524..6618
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     6535..6554
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    6572..6590
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6600..6618
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    6888..7005
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7054..7564
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7483..7501
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     intron          7573..8145
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     8153..8172
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        8200..8218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8229..8846
                     /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
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     primer_bind     complement(8302..8321)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gcgtacagcg gctcccggga gttctaggga tctgcccctc tccctccccc ccccctaacg
       61 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
      121 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
      181 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
      241 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
      301 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
      361 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
      421 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
      481 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
      541 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
      601 gatgataagg atccggaact aaaccatggc caagcctttg tctcaagaag aatccaccct
      661 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
      721 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
      781 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
      841 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
      901 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
      961 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     1021 ctaagcacaa tgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     1081 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     1141 atgcatcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     1201 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     1261 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     1321 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     1381 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1441 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     1501 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     1561 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1621 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     1681 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1741 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     1801 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     1861 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1921 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1981 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     2041 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2101 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2161 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2221 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2281 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2341 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2401 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2461 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2521 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2581 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2641 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2701 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2761 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2821 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2881 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2941 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3001 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3061 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3121 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3181 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3241 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3301 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3361 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3421 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3481 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3541 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3601 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3661 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3721 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3781 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3841 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3901 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3961 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4021 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4081 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4141 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4201 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4261 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4321 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4381 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4441 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4501 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     4561 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     4621 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     4681 agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt
     4741 aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt
     4801 ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat
     4861 tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata
     4921 cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     4981 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5041 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5101 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     5161 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     5221 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     5281 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     5341 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     5401 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     5461 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     5521 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     5581 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     5641 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     5701 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     5761 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     5821 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     5881 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     5941 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     6001 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     6061 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     6121 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     6181 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     6241 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     6301 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     6361 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     6421 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     6481 atctcgacgg tatcgatcac gagactagcc tcgagcggcc gcaatatttg catgtcgcta
     6541 tgtgttctgg gaaatcacca taaacgtgaa atccctatca gtgatagaga cttataagtt
     6601 ccctatcagt gatagagacg ctagcgtgtt gtaaatgagc acacaaaata cacatgctaa
     6661 aatattatat tctatgacct ttataaaatc aaccaaaatc ttctttttaa taactttagt
     6721 atcaataatt agaattttta tgttcctttt tgcaaacttt taataaaaat gagcaaaata
     6781 aaaaaacgct agttttagta actcgcgttg ttttcttcac ctttaataat agctactcca
     6841 ccacttgaat tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa
     6901 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     6961 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     7021 gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg
     7081 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     7141 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     7201 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     7261 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     7321 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     7381 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     7441 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     7501 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     7561 ccagggggat ctgtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa
     7621 ttcatgttat atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc
     7681 ttgtatcacc atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa
     7741 ccattgtctc ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg
     7801 catttgtaac gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc
     7861 taatcacttt tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag
     7921 agaacaattg ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg
     7981 cgtggaaata ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct
     8041 ttatggttac aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg
     8101 ggcccctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     8161 ctggttattg tgctgtctca tcattttggc aaagaattgt aatacgactc actatagggc
     8221 gaattgatat gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta
     8281 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     8341 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     8401 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     8461 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     8521 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     8581 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg
     8641 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     8701 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     8761 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     8821 aaaaacaact taaatgtgaa agtgggtcc
//
