LOCUS Exported 9488 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Twist-siRNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9488) AUTHORS Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C, Savagner P, Gitelman I, Richardson A, Weinberg RA TITLE Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. JOURNAL Cell 2004 Jun 25;117(7):927-39. PUBMED 15210113 REFERENCE 2 (bases 1 to 9488) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9488 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(605..1465) /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 1228..1247 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1466..1570) /gene="bla" /label=AmpR promoter primer_bind 1638..1656 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(1694..1716) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1816..1835 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2049..2068 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 2358..2376 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2358..2375 /label=SP6 /note="SP6 promoter, forward primer" LTR 2401..2634 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" misc_feature 2727..3315 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 2833..2844 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 3242..3262 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3386..3985) /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(3469..3489) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 3966..3985 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(3995..4324) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4009..4144 /label=SV40 ori /note="SV40 origin of replication" promoter 4356..4596 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 4356..4376 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 4527..4546 /label=LKO.1 5' /note="Human U6 promoter, forward primer" CDS complement(4923..4967) /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" misc_feature 5152..5385 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 5878..6003 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 6050..6683 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" polyA_signal 6851..6985 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6875..6894) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6929..6948 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(7410..7430) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" intron 7560..7625 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" promoter complement(8759..9088) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8773..8908 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(9163..9180) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9334..434) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9414..9433) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 61 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 1021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 1081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 1141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 1201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 1261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 1321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 1381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 1441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 1501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 1561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 1621 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 1681 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 1741 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 1801 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct 1861 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc 1921 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg 1981 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct 2041 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg 2101 ccggccacga tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg 2161 ctgaccattt ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa 2221 ccgactctga cggcagttta cgagagagat gatagggtct gcttcagtaa gccagatgct 2281 acacaattag gcttgtacat attgtcgtta gaacgcggct acaattaata cataacctta 2341 tgtatcatac acatacgatt taggtgacac tatagaatac acggaattaa ttctagatgc 2401 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac 2461 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca 2521 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc 2581 cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc 2641 ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag 2701 gtaccgagct cgaattaatt cccgatgcgg ggaggcggcc caaagggaga tccgactcgt 2761 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg 2821 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca 2881 ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca acaccacgga 2941 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag 3001 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt 3061 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg 3121 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga 3181 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc 3241 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta 3301 atccagaggt tgattatcgg aattaattcg atgcatgggg tcgtgcgctc ctttcggtcg 3361 ggcgctgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg caccaggtgc 3421 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg 3481 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct 3541 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc 3601 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca 3661 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg 3721 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt 3781 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca 3841 gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt 3901 cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc 3961 gcaccgtggg cttgtactcg gtcatggtaa gctttttgca aaagcctagg cctccaaaaa 4021 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa 4081 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg 4141 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca 4201 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat 4261 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac 4321 acacattcca cagggtcgac aaggtcgggc aggaagaggg cctatttccc atgattcctt 4381 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa 4441 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg 4501 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt 4561 tcgatttctt ggctttatat atcttgtgga aaggacgatt cgaaaagctg agcaagattc 4621 agaccttcaa gagaggtctg aatcttgctc agcttttgga tccgataccg tcgagatccg 4681 ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg 4741 ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc 4801 tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct 4861 cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc 4921 cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt 4981 tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac 5041 tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc 5101 caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt 5161 tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag 5221 actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc 5281 tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc 5341 atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt 5401 ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt 5461 tttactactt tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca 5521 ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc 5581 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag 5641 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg 5701 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc 5761 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc 5821 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac 5881 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac 5941 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga 6001 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt 6061 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg 6121 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct 6181 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca 6241 aaaagcagct gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc 6301 acgcctccct ggaaagtccc cagcggaaag tcccttgtag caagctcgat atcagcagtt 6361 cttgaagtac tccggatgca gctctcgggc cacgtgatga aatgctaggc ggctgtcaaa 6421 cctccactct aacacttctc tctccgggtc atccatccca tgcaggctca cagggtgtaa 6481 caagctggtg ttctctcctt tattggcctc ttctacctta tctggctcaa ctggtactag 6541 cttgtagcac catccaaagg tcagtggata tctgacccct ggccctggtg tgtagttctg 6601 ctaatcaggg aagtagcctt gtgtgtggta gatccacaga tcaaggatat cttgtcttct 6661 ttgggagtga attagccctt ccaactacta agtttgtagt acatatttaa caaatacaat 6721 ttctttaaaa tgaaaataat tcagaggaat cacaggttta gagtaaatga aaccacaggt 6781 aattggcagt ggtaataggg tatggggtgg gaagtttggg atgattttgg ttagcttgag 6841 ttatccagtt gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag 6901 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 6961 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 7021 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc 7081 tgattatgat ctctagtcaa ggcactatac atcaaatatt ccttattaac ccctttacaa 7141 attaaaaagc taaaggtaca caatttttga gcatagttat taatagcaga cactctatgc 7201 ctgtgtggag taagaaaaaa cagtatgtta tgattataac tgttatgcct acttataaag 7261 gttacagaat atttttccat aattttcttg tatagcagtg cagctttttc ctttgtggtg 7321 taaatagcaa agcaagcaag agttctatta ctaaacacag catgactcaa aaaacttagc 7381 aattctgaag gaaagtcctt ggggtcttct acctttctct tcttttttgg aggagtagaa 7441 tgttgagagt cagcagtagc ctcatcatca ctagatggca tttcttctga gcaaaacagg 7501 ttttcctcat taaaggcatt ccaccactgc tcccattcat cagttccata ggttggaatc 7561 taaaatacac aaacaattag aatcagtagt ttaacacatt atacacttaa aaattttata 7621 tttaccttag agctttaaat ctctgtaggt agtttgtcca attatgtcac accacagaag 7681 taaggttcct tcacaaagat ccggggccca ctcataaatc cagttgccgc cacggtagcc 7741 aatcaccgta tcgtataaat catcgtcggt acgttcggca tcgctcatca caatacgtgc 7801 ctggacgtcg aggatttcgc gtgggtcaat gccgcgccag atccacatca gacggttaat 7861 catgcgatac cagtgaggga tggttttacc atcaagggcc gactgcacag gcggttgtgc 7921 gccgtgatta aagcggcgga ctagcgtcga ggtttcagga tgtttaaagc ggggtttgaa 7981 cagggtttcg ctcaggtttg cctgtgtcat ggatgcagcc tccagaatac ttactggaaa 8041 ctattgtaac ccgcctgaag ttaaaaagaa caacgcccgg cagtgccagg cgttgaaaag 8101 attagcgacc ggagattggc gggacgaata cgacgcccat atcccacggc tgttcaatcc 8161 aggtatcttg cgggatatca acaacatagt catcaaccag cggacgacca gccggttttg 8221 cgaagatggt gacaaagtgc gcttttggat acatttcacg aatcgcaacc gcagtaccac 8281 cggtatccac caggtcatca ataacgatga agccttcgcc atcgccttct gcgcgtttca 8341 gcactttaag ctcgcgctgg ttgtcgtgat cgtagctgga aatacaaacg gtatcgacat 8401 gacgaatacc cagttcacgc gccagtaacg cacccggtac agaccgccac ggcttacggc 8461 aataatgcct ttccattgtt cagaaggcat cagtcggctt gcgagtttac gtgcatggat 8521 ctgcaacatg tcccaggtga cgatgtattt ttcgctcatg tgaagtgtcc cagcctgttt 8581 atctacggct taaaaagtgt tcgaggggaa aataggttgc gcgagattat agagatctgg 8641 cgcactaaaa accagtattt cacatgagtc cgcgtctttt tacgcactgc ctctccctga 8701 cgcgggataa agtggtattc tcaaacatat ctcgcaagcc tgtcttgtgt ccaagctttt 8761 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg 8821 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg 8881 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg 8941 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac 9001 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg 9061 gggactttcc acaccctaac tgacacacat tccacagctg cattaatgaa tcggccaacg 9121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 9181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 9241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 9301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 9361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 9421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 9481 cggatacc //