LOCUS       sequence-66372-e        8036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lentiviral vector encoding a fusion between codon-optimized 
            P.aeruginosa exoY mutant (K81M) and myc-tagged destabilized FKBP12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8036)
  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 8036)
  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: pMA3228
FEATURES             Location/Qualifiers
     source          1..8036
                     /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"
     CDS             4074..4103
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4104..4427
                     /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(4469..4489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4474..4803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4654..4789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4817..5416
                     /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(4817..4836)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5313..5333
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6023..6203
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6265..6853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6345..6364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7024..7815,7816..7884))
                     /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     7647..7666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7885..7989)
                     /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 cgccaccatg aggatcgacg
     2941 gccacaggca ggtggtgagc aatgccacag cccagcctgg ccccctgctg aggcctgctg
     3001 acatgcaggc cagggccctc caggacctgt ttgatgccca gggagtggga gtgcctgtgg
     3061 agcatgccct gaggatgcag gctgtggcca ggcagaccaa cacagtgttt ggcatcaggc
     3121 ctgtggagag gattgtgacc accctgattg aggagggctt ccccaccatg ggcttctctg
     3181 tgaagggcaa gagcagcaac tggggccccc aggctggctt catctgtgtg gaccagcacc
     3241 tgagcaagag ggaggacagg gacacagctg agatcaggaa gctgaacctg gctgtggcca
     3301 agggcatgga tggaggagcc tacacccaga cagacctgag gatcagcagg cagaggctgg
     3361 ctgagctggt gaggaacttt ggcctggtgg ctgatggagt gggccctgtg aggctgctga
     3421 cagcccaggg cccctctggc aagaggtatg agtttgaggc caggcaggag gctgatggcc
     3481 tgtacaggat cagcaggctg ggcaggagcg aggccgtgca ggtgctggcc agccccgcct
     3541 gcggcctggc catgaccgcc gactacgacc tgttcctggt ggcccccagc atcgaggccc
     3601 acggcagcgg cggcctggac gccaggagga acaccgccgt gaggtacacc cccctgggcg
     3661 ccaaggaccc cctgagcgag gacggcttct acggcaggga ggacatggcc aggggcaaca
     3721 tcacccccag gaccaggcag ctggtggacg ccctgaacga ctgcctgggc aggggcgagc
     3781 acagggagat gttccaccac agcgacgacg ccggcaaccc cggcagccac atgggcgaca
     3841 acttccccgc caccttctac ctgcccaggg cgatggagca cagggtgggc gaggagagcg
     3901 tgaggttcga cgaggtgtgc gtggtggccg acaggaagag cttcagcctg ctggtggagt
     3961 gcatcaaggg caacggctac cacttcaccg cccaccccga ctggaacgtg cccctgaggc
     4021 ccggcttcca ggaggccctg gacttcttcc agaggaaggt ggagctggtt ggggagcaga
     4081 aactcatcag cgaggaggac ctgggagtgc aggtggaaac catctcccca ggagacgggc
     4141 gcaccttccc caagcgcggc cagacctgtg tggtgcacta caccgggatg cttggagatg
     4201 gaaagaaagt cgattcctct cgggacagaa acaagccctt taagtttatg ctaggcaagc
     4261 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagggagcca
     4321 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     4381 cacatgccac tctcgtcttc gatgtggagc ttctggaact ggaataaatg caggaattga
     4441 tggccgccac cgcggtggag gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4741 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4801 aaaaagctag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4861 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4921 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4981 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5041 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5101 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5161 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5221 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5281 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5341 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5401 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     5461 accccatgca tcgataccgt caattcgatt aagtcgaggg tcgatcgacc tcgatcgaga
     5521 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     5581 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     5641 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     5701 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     5761 cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc
     5821 actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc
     5881 caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5941 ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg
     6001 agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6061 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6121 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6181 gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6241 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6301 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6601 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6661 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6721 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6781 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6841 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6901 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6961 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7021 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7081 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7201 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7441 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7681 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7861 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7921 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7981 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gatcgacgga tcggga
//
