LOCUS       sequence-66412-e        6990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for the dual control of expression through Tet-On 
            promoter and C-terminal destabilizing domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Ochoa CD, Alexeyev M, Pastukh V, Balczon R, Stevens T
  TITLE     Pseudomonas aeruginosa exotoxin Y is a promiscuous cyclase that 
            increases endothelial tau phosphorylation and permeability.
  JOURNAL   J Biol Chem. 2012 Jul 20;287(30):25407-18. doi: 
            10.1074/jbc.M111.301440. Epub 2012 May 25.
  PUBMED    22637478
REFERENCE   2  (bases 1 to 6990)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M111.301440"; journalName: "J Biol Chem"; date: 
            "2012-07-20- 20"; volume: "287"; issue: "30"; pages: "25407-18"
COMMENT     Sequence Label: pMA3174
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        227..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1016..1141
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1638..1871
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2056..2100
                     /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    2394..2511
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2597..2911
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2605..2623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2641..2659
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2676..2694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2712..2730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2748..2766
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2783..2801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2819..2837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2884..2908
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2936..2952)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2937..2953)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             3028..3057
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3058..3381
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD-C
                     /note="FKBP12 mutant optimized for use as a C-terminal tag"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLGDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQGAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     ELE"
     primer_bind     complement(3423..3443)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3428..3757
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3608..3743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3771..4370
                     /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(3771..3790)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4267..4287
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4977..5157
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5219..5807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5299..5318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5978..6769,6770..6838))
                     /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     6601..6620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6839..6943)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg cgcgttgaca ttgattattg
      241 actagctagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atagaccaga tctgagcctg ggagctctct ggctaactag
      841 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      901 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      961 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     1021 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     1081 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     1141 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     1201 aaagaaaaaa tataaataaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1261 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1321 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1381 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1441 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgcgct gatcttcaga
     1501 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1561 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1621 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1681 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1741 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1801 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1861 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1921 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     1981 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     2041 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     2101 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     2161 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     2221 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2281 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2341 aacggatcgg cactgcgtgc gccaattcga caaatggcag tattcatcca caattttaaa
     2401 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     2461 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     2521 tacagggaca gcagagatcc agtttggtta gtaccgggcc ggcccgctct agctagacgt
     2581 attaccgcca gacctcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2641 tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat
     2701 gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
     2761 tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc
     2821 cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag
     2881 cagagctcgt ttagtgaacc gtcagatcgc ctggagaatt cgatatcaag cttatcgata
     2941 ccgtcgacct cgaggggggg cccggtaccg aggacgcgtc aattcgagct ccaccgcggt
     3001 ggcggccgct ctagaactag tcccggggag cagaaactca tcagcgagga ggacctggga
     3061 gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc
     3121 tgtgtggtgc actacaccgg gatgcttgga gatggaaaga aagtcgattc ctctcgggac
     3181 agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa
     3241 ggggttgccc agatgagtgt gggtcaggga gccaaactga ctatatctcc agattatgcc
     3301 tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg
     3361 gagcttctgg aactggaata aatgcaggaa ttgatggccg ccaccgcggt ggaggaccct
     3421 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3481 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3601 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3661 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3721 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctagcttacc atgaccgagt
     3781 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3841 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     3901 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     3961 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4021 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4081 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4141 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4201 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4261 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4321 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     4381 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgata ccgtcaattc
     4441 gattaagtcg agggtcgatc gacctcgatc gagacctaga aaaacatgga gcaatcacaa
     4501 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     4561 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     4621 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4681 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     4741 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     4801 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     4861 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     4921 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     4981 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5041 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5101 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
     5161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6961 tgccacctga cgtcgatcga cggatcggga
//
