EZ-Tet-pLKO-Hygro

LOCUS       EZ-Tet-pLKO-Hygro                9469 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Tet/Dox inducible shRNA lentivirus with hygromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    EZ-Tet-pLKO-Hygro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9469)
  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 9469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9469
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             120..164
                     /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        453..547
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     464..483
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    501..519
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    529..547
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    817..934
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        983..1493
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1412..1430
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     intron          1502..2074
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        2128..2146
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2157..2774
                     /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(2230..2249)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    2812..3385
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2979..2996)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3206..3225
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3392..4429
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4408..4416
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     LTR             4617..4850
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4922..5043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4959..4978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5013..5032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5083..5218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5145..5164
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        complement(5239..5257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5267..5284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5267..5283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5276..5298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5425..5880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5512..5531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5722..5743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5906..6010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6011..6871
                     /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(6229..6248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7042..7630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7531..7550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7784..7801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7918..7939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7954..7984
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7992..8008
                     /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     7997..8019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8016..8032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8016..8032
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8051..8071
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8053..8071
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8099..8325
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8326..8506
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8553..8678
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9171..9404
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
       61 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
      121 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
      181 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
      241 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      301 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
      361 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
      421 atcgatcacg agactagcct cgagcggccg caatatttgc atgtcgctat gtgttctggg
      481 aaatcaccat aaacgtgaaa tccctatcag tgatagagac ttataagttc cctatcagtg
      541 atagagacgc tagcgtgttg taaatgagca cacaaaatac acatgctaaa atattatatt
      601 ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta tcaataatta
      661 gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa aaaaacgcta
      721 gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac cacttgaatt
      781 ctcgacctcg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
      841 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
      901 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
      961 tccactttgg ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt
     1021 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     1081 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     1141 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     1201 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     1261 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     1321 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     1381 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     1441 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     1501 tgtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata
     1561 tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca
     1621 tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc
     1681 tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg
     1741 aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt
     1801 ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt
     1861 tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat
     1921 tcttattggt agaaacaact acatcctggt catcatcctg cctttctctt tatggttaca
     1981 atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
     2041 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacggct ggttattgtg
     2101 ctgtctcatc attttggcaa agaattgtaa tacgactcac tatagggcga attgatatgt
     2161 ctagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
     2221 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
     2281 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
     2341 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     2401 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     2461 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     2521 aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt
     2581 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     2641 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     2701 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     2761 aatgtgaaag tgggtccgcg tacagcggct cccgggagtt ctagggatct gcccctctcc
     2821 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2881 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2941 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     3001 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     3061 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     3121 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3181 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     3241 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3301 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     3361 tttcctttga aaaacacgat gataaggatc catgaagaag cccgaactca ccgctaccag
     3421 cgttgaaaaa tttctcatcg agaagttcga cagtgtgagc gacctgatgc agttgtcgga
     3481 gggcgaagag agccgagcct tcagcttcga tgtcggcgga cgcggctatg tactgcgggt
     3541 gaatagctgc gctgatggct tctacaaaga ccgctacgtg taccgccact tcgccagcgc
     3601 tgcactaccc atccccgaag tgttggacat cggcgagttc agcgagagcc tgacatactg
     3661 catcagtaga cgcgcccaag gcgttactct ccaagacctc cccgaaacag agctgcctgc
     3721 tgtgttacag cctgtcgccg aagctatgga tgctattgcc gccgccgacc tcagtcaaac
     3781 cagcggcttc ggcccattcg ggccccaagg catcggccag tacacaacct ggcgggattt
     3841 catttgcgcc attgctgatc cccatgtcta ccactggcag accgtgatgg acgacaccgt
     3901 gtccgccagc gtagctcaag ccctggacga actgatgctg tgggccgaag actgtcccga
     3961 ggtgcgccac ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg acaacggccg
     4021 catcaccgcc gtaatcgact ggtccgaagc tatgttcggg gacagtcagt acgaggtggc
     4081 caacatcttc ttctggcggc cctggctggc ttgcatggag cagcagactc gctacttcga
     4141 gcgccggcat cccgagctgg ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg
     4201 cctggatcag ctctaccaga gcctcgtgga cggcaacttc gacgatgctg cctgggctca
     4261 aggccgctgc gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc
     4321 tcgccggagc gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa
     4381 ccgccggccc agtacacgac cgcgcgctaa ggaggtaggt cgagtttaag acgtcgaggt
     4441 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     4501 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcggtac ctttaagacc
     4561 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4621 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     4681 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4741 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4801 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     4861 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     4921 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4981 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5041 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     5281 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5341 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5401 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5641 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5701 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5761 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5821 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6001 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6061 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7861 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7921 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7981 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     8041 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     8101 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     8161 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     8221 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     8281 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     8341 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8401 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8461 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     8521 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     8581 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     8641 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     8701 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     8761 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     8821 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     8881 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     8941 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     9001 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     9061 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     9121 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     9181 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     9241 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     9301 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     9361 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     9421 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttgga
//