LOCUS Exported 7084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To knockdown Thioredoxin-1. ACCESSION . VERSION . KEYWORDS plko-puromycin-shTRX1-259 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) AUTHORS Young JJ, Patel A, Rai P TITLE Suppression of thioredoxin-1 induces premature senescence in normal human fibroblasts. JOURNAL Biochem Biophys Res Commun. 2010 Feb 12;392(3):363-8. doi: 10.1016/j.bbrc.2010.01.026. Epub 2010 Jan 13. PUBMED 20074557 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" protein_bind 1..22 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 37..67 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 75..91 /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 80..102 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 99..115 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 99..115 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 134..154 /label=T3 /note="T3 promoter, forward primer" promoter 136..154 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 182..408 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 409..589 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 636..761 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1254..1487 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1672..1716 /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 2014..2254 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2014..2034 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2185..2204 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 2359..2476 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2525..3035 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 2954..2972 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 3057..3656 /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(3057..3076) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3553..3573 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3784..4017 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4089..4210 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4126..4145) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4180..4199 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4250..4385 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4312..4331 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4405..4424) /label=T7 /note="T7 promoter, forward primer" promoter complement(4406..4424) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4434..4451) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4434..4450) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4443..4465) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4592..5047 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4679..4698) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4889..4910 /label=F1ori-F /note="F1 origin, forward primer" promoter 5073..5177 /gene="bla" /label=AmpR promoter CDS 5178..6038 /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(5396..5415) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6209..6797 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6698..6717 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6951..6968 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta 181 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat 241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat 301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg 361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg 421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga 601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg 661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga 721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa 781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa 841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag 901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat 961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga 1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta 1081 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac 1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca 1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct 1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg 1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg 1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag 1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt 1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa 1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat 1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa 1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat 1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata 1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt 1861 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt 1921 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca 1981 cgagactagc ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca 2041 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac 2101 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca 2161 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc 2221 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggcttca gagtgtgaag 2281 tcaaactcga gtttgacttc acactctgaa gctttttgaa ttctcgacct cgagacaaat 2341 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga 2401 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac 2461 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggccgcggct 2521 cgagggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct 2581 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca 2641 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc 2701 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac 2761 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg 2821 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga 2881 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt 2941 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct 3001 ccctcgttga ccgaatcacc gacctctctc cccaggggga tccaccggag cttaccatga 3061 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca 3121 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc 3181 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 3241 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 3301 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 3361 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 3421 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 3481 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 3541 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 3601 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc 3661 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcggtacctt 3721 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 3781 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt 3841 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 3901 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 3961 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 4021 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 4081 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4141 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4201 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct 4261 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4321 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 4381 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc 4441 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 4501 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 4561 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg 4621 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 4681 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 4741 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 4801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4921 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 4981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt 5041 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 5101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 5161 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 5221 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 5281 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 5341 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 5401 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 5461 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 5521 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 5581 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 5641 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 5701 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 5761 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 5821 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 5881 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 5941 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 6001 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 6061 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 6121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 6181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 6241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 6301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 6361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 6421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 6481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 6541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 6601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 6661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 6721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 6781 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 6841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 6901 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 6961 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 7021 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 7081 caat //