LOCUS Exported 7979 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral plasmid expressing the reverse tetracycline transactivator. ACCESSION . VERSION . KEYWORDS FUW-M2rtTA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7979) AUTHORS Hockemeyer D, Soldner F, Cook EG, Gao Q, Mitalipova M, Jaenisch R TITLE A drug-inducible system for direct reprogramming of human somatic cells to pluripotency. JOURNAL Cell Stem Cell. 2008 Sep 11. 3(3):346-53. PUBMED 18786421 REFERENCE 2 (bases 1 to 7979) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7979 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 618..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 834..1014 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1061..1186 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1679..1912 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2097..2141 /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 2439..2556 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2621..3832 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3669..3688 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3861..4607 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=rtTA-Advanced /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" misc_feature 4644..5232 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4697..4717) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5115..5126) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5235..5251) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5236..5252) /label=pBluescriptKS /note="For pBluescript vector" LTR 5757..5937 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5963..5980) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5969..6080 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(6226..6814) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6306..6325) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6985..7845) /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 7608..7627 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7846..7950) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc 841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga 1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag 1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag 1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga 1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc 1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc 1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt 1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta 1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta 2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga 2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa 2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa 2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat 2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag 2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg 2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt 2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa 2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga 2581 gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg 2641 gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc 2701 agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac 2761 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct 2821 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc 2881 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg 2941 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat 3001 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg 3061 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc 3121 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc 3181 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg 3241 gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct 3301 tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga 3361 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca 3421 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt 3481 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg 3541 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg 3601 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct 3661 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga 3721 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta 3781 gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg 3841 ggcccgggaa ttaattcacc atgtctagac tggacaagag caaagtcata aacggcgctc 3901 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc 3961 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg 4021 ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt 4081 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc 4141 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg 4201 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc 4261 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag 4321 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa 4381 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca 4441 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg 4501 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc 4561 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta agtaaggatc 4621 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga 4681 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 4741 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 4801 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 4861 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 4921 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 4981 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat 5041 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 5101 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 5161 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 5221 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa 5281 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg 5341 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg 5401 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac 5461 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc 5521 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc 5581 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt 5641 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt 5701 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt 5761 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5821 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5881 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg 5941 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 6001 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 6061 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 6121 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgt gagcaaaagg 6181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 7021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 7081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 7141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 7201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 7261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 7321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 7381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 7441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 7501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 7561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 7621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 7681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7921 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg //