LOCUS       sequence-66377-e        8700 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A lentiviral vector for the doxycycline-inducible expression of 
            soluble guanylate cyclase1 alpha3 mutant R592Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8700)
  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 8700)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pMA3431
FEATURES             Location/Qualifiers
     source          1..8700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1035..1160
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1657..1890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2075..2119
                     /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    2413..2530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2616..2930
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2624..2642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2660..2678
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2695..2713
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2731..2749
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2767..2785
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2802..2820
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2838..2856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2903..2927
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5066..5095
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5152..5172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5157..5486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5337..5472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5500..6099
                     /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(5500..5519)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5996..6016
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6706..6886
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6948..7536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7028..7047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7707..8498,8499..8567))
                     /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     8330..8349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8568..8672)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
      241 gcgttgacat tgattattga ctagctagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tagaccagat ctgagcctgg
      841 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      901 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      961 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     1021 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1081 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1141 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1201 ttcggttaag gccaggggga aagaaaaaat ataaataaac atatagtatg ggcaagcagg
     1261 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     1321 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1381 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1441 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1501 ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1561 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1621 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1681 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1741 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1801 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1861 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1921 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1981 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     2041 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     2101 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     2161 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2221 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2281 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2341 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattcgac aaatggcagt
     2401 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2461 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2521 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag taccgggccg
     2581 gcccgctcta gctagacgta ttaccgccag acctcgagtt tactccctat cagtgataga
     2641 gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt ttactcccta
     2701 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
     2761 gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca gtgatagaga
     2821 acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac
     2881 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagaattc
     2941 gatatcaagc ttatcgatac cgtcgacggt atcgataagc ttgatgcgtg ccaccatgtt
     3001 ctgcaggaag ttcaaagatc tcaagatcac gggggagtgt cctttctcct tactggctcc
     3061 tggtcaggtt cctacggagc caatagagga ggtggccgga gtctctgaga gctgccaggc
     3121 cactctgccc acctgccagg agtttgctga gaatgcagaa gggagtcacc cccaaagaaa
     3181 gacaagccgc aacagagtct acctacacac cctggcagag agtattggca agctcatttt
     3241 cccagagttt gagagactga accttgcact tcagagaaca ttggcaaagc acaaaataaa
     3301 ggaaaacagg aattcttcag aaaaagaaga ccttgaaaga ataattgcag aagaagcaat
     3361 tgcggcaggt gtcccagtgg aggcgctcaa agactctctg ggtgaggaac tgttcaagat
     3421 ctgttatgag gaagatgagc acatcctggg cgtggttggc ggcaccctga aggactttct
     3481 caacagcttc agcacgctcc tgaagcagag cagccactgc caggaggcgg agaggcgggg
     3541 gcgactggaa gatgcctcca tcttgtgcct ggacaaggac caggactttc taaatgttta
     3601 ctacttcttc ccgaagagaa ccacagccct gcttctcccc ggtatcatta aagcggctgc
     3661 tcgcatactg tacgaaagcc acgtggaggt gtccctgatg cctccctgct tccgaagtga
     3721 gtgtaccgaa ttcgtgaacc agccctattt gctctactcg gtccacgtga agagcaccaa
     3781 gccttctctg tccccaggca aaccccagtc ctcgctagtg atccctactt cgctcttctg
     3841 caaaactttc ccattccact tcatgctgga ccgtgacctg gccattctgc agctgggtaa
     3901 tggcatcaga aggctggtga acaagaggga cttccaaggg aagcccaact ttgaagagtt
     3961 ctttgaaatt ctaactccca aaatcaacca gacatttagc ggcatcatga caatgttgaa
     4021 tatgcagttt gtcatacggg tgaggagatg ggataactta gtgaagaaat cttcaagggt
     4081 tatggacctc aaaggtcaaa tgatctacat cgttgaatcc agtgccatct tgttcttagg
     4141 gtcgccgtgt gtggacagat tggaagattt cacaggacgg ggtctctatc tgtctgacat
     4201 cccgattcat aatgccctga gggatgttgt cttgataggg gaacaggcac gggctcaaga
     4261 tggcctcaag aagaggttgg gaaagctgaa ggcaaccttg gagcatgccc accaagccct
     4321 ggaggaggag aagaagaaga cggtagatct cctgtgttct atctttccct ctgaggttgc
     4381 tcagcagctc tggcaaggac aaattgtgca agccaagaag ttcaatgagg tcaccatgct
     4441 cttctcagat atcgtagggt tcactgccat ctgctctcag tgttcacctc tgcaggtcat
     4501 caccatgctc aacgctctct atactcgctt tgaccaacag tgtggagagc tggatgtcta
     4561 caaggtggag accatcggag atgcatattg tgtggcaggg ggattgcaca gagagagtga
     4621 cacacatgct gtccagatag cactgatggc cctgaagatg atggagctct ccaatgaggt
     4681 catgtctccc cacggagaac ctatcaagat gcgaatcgga ctgcattctg ggtcagtgtt
     4741 tgctggagtt gttggagtga agatgcccca gtattgcctg tttggaaaca atgtcactct
     4801 ggctaacaaa tttgagtcct gcagtgtgcc tcggaaaatc aatgtcagcc ccacgacata
     4861 caggttactc aaagactgtc ctggctttgt gttcaccccg agatcgaggg aggagcttcc
     4921 accaaacttc cctagtgaca ttcctggaat ctgtcacttt ctggatgctt atcaacatca
     4981 aggacctaat tctaaaccat ggttccagca gaaagatgca gaagatggaa acgctaactt
     5041 cttaggcaaa gcatcagggg tagatgagca gaaactcatc agcgaggagg acctgtgagg
     5101 atcccccggg ctgcaggaat tgatggccgc caccgcggtg gaggaccctg tggaatgtgt
     5161 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5281 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5401 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5461 tttttggagg cctaggcttt tgcaaaaagc tagcttacca tgaccgagta caagcccacg
     5521 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     5581 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     5641 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5701 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5761 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5821 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5881 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5941 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6001 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     6061 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     6121 cgcccgaccg aaaggagcgc acgaccccat gcatcgatac cgtcaattcg attaagtcga
     6181 gggtcgatcg acctcgatcg agacctagaa aaacatggag caatcacaag tagcaataca
     6241 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6301 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6361 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     6421 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
     6481 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     6541 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg
     6601 agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc
     6661 ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta
     6721 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6781 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6841 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa
     6901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
