LOCUS Exported 7084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION shETV5 shRNA. ACCESSION . VERSION . KEYWORDS pLKO.1 shETV5-B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) AUTHORS Wang B, Krall EB, Aguirre AJ, Kim M, Widlund HR, Doshi MB, Sicinska E, Sulahian R, Goodale A, Cowley GS, Piccioni F, Doench JG, Root DE, Hahn WC TITLE ATXN1L, CIC, and ETS Transcription Factors Modulate Sensitivity to MAPK Pathway Inhibition. JOURNAL Cell Rep. 2017 Feb 7;18(6):1543-1557. doi: 10.1016/j.celrep.2017.01.031. PUBMED 28178529 REFERENCE 2 (bases 1 to 7084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7084 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 41..158 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 217..717 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 636..654 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 739..1338 /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(739..758) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1235..1255 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 1466..1699 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 1771..1892 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1808..1827) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1862..1881 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 1932..2067 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1994..2013 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2087..2106) /label=T7 /note="T7 promoter, forward primer" promoter complement(2088..2106) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2116..2133) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2116..2132) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2125..2147) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2274..2729 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2361..2380) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2571..2592 /label=F1ori-F /note="F1 origin, forward primer" promoter 2755..2859 /gene="bla" /label=AmpR promoter CDS 2860..3720 /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(3078..3097) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3891..4479 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4380..4399 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4633..4650 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4767..4788 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4803..4833 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4841..4857 /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 4846..4868 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4865..4881 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4865..4881 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 4900..4920 /label=T3 /note="T3 promoter, forward primer" promoter 4902..4920 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 4948..5174 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5175..5355 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5402..5527 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6020..6253 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6438..6482 /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 6780..7020 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 6780..6800 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 6951..6970 /label=LKO.1 5' /note="Human U6 promoter, forward primer" ORIGIN 1 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga 61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca 181 gagatccact ttggcgccgg ctcgaggggg cccgggttgc gccttttcca aggcagccct 241 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac 301 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc 361 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg 421 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac 481 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc 541 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt 601 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg 661 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg 721 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac 781 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc 841 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc 901 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg 961 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc 1021 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg 1081 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac 1141 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc 1201 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc 1261 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc 1321 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca 1381 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 1441 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat 1501 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 1561 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 1621 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 1681 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt 1741 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac 1801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 1861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc 1921 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 1981 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 2041 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc 2101 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc 2281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2401 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2641 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2701 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 2761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 2821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 2881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 2941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 3001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 3061 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 3121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 3181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 3241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 3301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 3361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 3421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 3481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 3541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 3601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 3661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 3721 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 3781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 3841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 3901 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 3961 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 4021 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 4081 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 4141 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4201 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4261 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4321 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 4381 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 4441 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 4501 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 4561 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 4621 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 4681 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 4741 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 4801 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 4861 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg 4921 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca 4981 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc 5041 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga 5101 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag 5161 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct 5221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 5281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 5341 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga 5401 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg 5461 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga 5521 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc 5581 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg 5641 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca 5701 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc 5761 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa 5821 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca 5881 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag 5941 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag 6001 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca 6061 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag 6121 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca 6181 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg 6241 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc 6301 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga 6361 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat 6421 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt 6481 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag 6541 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta 6601 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca 6661 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag 6721 tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg 6781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga 6841 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa 6901 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata 6961 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac 7021 gaggtaccgg ccgtgacact tagtacatta actcgagtta atgtactaag tgtcacggtt 7081 tttg //