LOCUS Exported 8394 bp ds-DNA circular SYN 10-FEB-2022 DEFINITION 3rd gen lentiviral reverse tetracycline-controlled transactivator 3 (rtTA3) expression vector, CMV promoter, Blast. ACCESSION . VERSION . KEYWORDS pLenti CMV rtTA3 Blast (w756-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8394) TITLE Tet-On Advanced vectors REFERENCE 2 (bases 1 to 8394) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 10, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8394 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 98..115 /label=L4440 /note="L4440 vector, forward primer" protein_bind 232..253 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 268..298 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 306..322 /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 311..333 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 330..346 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 330..346 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 365..385 /label=T3 /note="T3 promoter, forward primer" promoter 367..385 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 413..639 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 640..820 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 867..992 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1485..1718 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1903..1947 /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 2214..2330 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2353..2656 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2657..2860 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2811..2831 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2857..2881 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 2965..2989 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2992..3699 /codon_start=1 /product="reverse tetracycline transactivator 3" /label=rtTA3 /note="activates expression from Tet-responsive promoters in the presence of doxycycline" /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPG" protein_bind complement(3698..3722) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 3753..4341 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3806..3826) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4224..4235) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4392..4412) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4397..4726 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4577..4712 /label=SV40 ori /note="SV40 origin of replication" promoter 4774..4821 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4840..5238 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 5325..5558 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5630..5751 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5667..5686) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5721..5740 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5791..5926 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5946..5965) /label=T7 /note="T7 promoter, forward primer" promoter complement(5947..5965) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5975..5992) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5975..5991) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5984..6006) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6133..6588 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6220..6239) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6430..6451 /label=F1ori-F /note="F1 origin, forward primer" promoter 6614..6718 /gene="bla" /label=AmpR promoter CDS 6719..7579 /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(6937..6956) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7750..8338 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8239..8258 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa 361 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct 421 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa 481 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta 541 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc 601 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag 661 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 721 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 781 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact 841 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc 901 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag 961 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc 1021 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag 1081 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag 1141 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac 1201 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa 1261 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg 1321 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga 1381 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag 1441 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt 1501 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag 1561 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag 1621 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc 1681 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga 1741 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa 1801 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc 1861 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta 1921 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg 1981 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct 2041 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac 2101 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga 2161 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa 2221 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca 2281 gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt atcgataagc 2341 ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 2401 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 2461 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 2521 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 2581 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 2641 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 2701 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 2761 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 2821 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat 2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccgac tctagtccag 2941 tgtggtggaa ttctgcagat atcaacaagt ttgtacaaaa aagttggcac catgtctaga 3001 ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa 3061 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg 3121 cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat 3181 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc 3241 aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc 3301 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa 3361 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc 3421 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc 3481 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa 3541 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc 3601 gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat 3661 gcccttgacg attttgacct tgacatgctc cccgggtaat caactttctt gtacaaagtg 3721 gttgatatcc agcacagtgg cggccgctcg acaatcaacc tctggattac aaaatttgtg 3781 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 3841 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 3901 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 3961 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 4021 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 4081 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 4141 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg 4201 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 4261 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 4321 ccctttgggc cgcctccccg cctggaattc tgcagatatc cggttagtaa tgagtttgga 4381 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4441 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 4501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 4561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 4681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 4741 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat 4801 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa 4861 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa 4921 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat 4981 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct 5041 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg 5101 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata 5161 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt 5221 tatgtgtggg agggctaagc acaattcgag ctcggtacct ttaagaccaa tgacttacaa 5281 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctagctca 5341 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 5401 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 5461 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 5521 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 5581 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 5641 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5701 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5761 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 5821 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 5881 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca 5941 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg 6001 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca 6061 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga 6121 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 6181 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 6241 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 6301 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 6361 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 6421 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 6481 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 6541 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt 6601 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 6661 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 6721 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 6781 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 6841 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 6901 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 6961 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 7021 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 7081 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 7141 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 7201 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 7261 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 7321 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 7381 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 7441 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 7501 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 7561 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 7621 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 7681 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 7741 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 7801 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 7861 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 7921 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 7981 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 8041 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 8101 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 8161 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 8221 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 8281 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 8341 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctca //