LOCUS       sequence-90361-e        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral pBabe puro vector encoding N-terminally FLAG tagged 
            mutant human IRE1a deleted from amino acids P28 to D408 and bearing 
            mutation K599A (kinase domain mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  AUTHORS   Volmer R, van der Ploeg K, Ron D
  TITLE     Membrane lipid saturation activates endoplasmic reticulum unfolded 
            protein response transducers through their transmembrane domains.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Mar 19;110(12):4628-33. doi: 
            10.1073/pnas.1217611110. Epub 2013 Mar 4.
  PUBMED    23487760
REFERENCE   2  (bases 1 to 6980)
  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: 
            "10.1073/pnas.1217611110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-03-19- 19"; volume: "110"; issue: "12"; pages: "4628-33"
COMMENT     Sequence Label: Flag_HsIRE1a_deltaP29_D408_K599A_pBabePuro
FEATURES             Location/Qualifiers
     source          1..6980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    523..722
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             923..1339
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1269..1291
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1307..1323
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1343..1387
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1388..1411
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3291..3311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3296..3625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3476..3611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3635..4234
                     /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(3635..3654)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4131..4151
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4455..4924
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5054..5071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5225..5813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5305..5324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5973..6764,6765..6833))
                     /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     6596..6615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6834..6938)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6969..6980,1..458)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
       61 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      121 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
      181 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
      241 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
      301 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
      361 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      421 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
      481 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
      541 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
      601 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      661 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
      721 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
      781 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
      841 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      901 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
      961 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1021 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1081 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1141 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1201 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1261 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1321 cactccttct ctaggcgccg gcatgtctgc acttctgatc ctagctcttg ttggagctgc
     1381 agttgctgac tacaaagacg atgacgacaa gctgattttt ggaagtacca gcacagtgac
     1441 gcttcagact tcagaaaacg cacctaccac cgtgtctcgg gatgtggagg agaagcccgc
     1501 ccatgcccct gcccggcccg aggcccccgt ggactccatg cttaaggaca tggctaccat
     1561 catcctgagc accttcctgc tgattggctg ggtggccttc atcatcacct atcccctgag
     1621 catgcatcag cagcagcagc tccagcacca gcagttccag aaggaactgg agaagatcca
     1681 gctcctgcag cagcagcagc agcagctgcc cttccaccca cctggagaca cggctcagga
     1741 cggcgagctc ctggacacgt ctggcccgta ctcagagagc tcgggcacca gcagccccag
     1801 cacgtccccc agggcctcca accactcgct ctgctccggc agctctgcct ccaaggctgg
     1861 cagcagcccc tccctggaac aagacgatgg agatgaggaa accagcgtgg tgatagttgg
     1921 gaaaatttcc ttctgtccca aggatgtcct gggccatgga gctgagggca caattgtgta
     1981 ccggggcatg tttgacaacc gcgacgtggc cgtggcgagg atcctccccg agtgttttag
     2041 cttcgcagac cgtgaggtcc agctgttgcg agaatcggat gagcacccga acgtgatccg
     2101 ctacttctgc acggagaagg accggcaatt ccagtacatt gccatcgagc tgtgtgcagc
     2161 caccctgcaa gagtatgtgg agcagaagga ctttgcgcat ctcggcctgg agcccatcac
     2221 cttgctgcag cagaccacct cgggcctggc ccacctccac tccctcaaca tcgttcacag
     2281 agacctaaag ccacacaaca tcctcatatc catgcccaat gcacacggca agatcaaggc
     2341 catgatctcc gactttggcc tctgcaagaa gctggcagtg ggcagacaca gtttcagccg
     2401 ccgatctggg gtgcctggca cagaaggctg gatcgctcca gagatgctga gcgaagactg
     2461 taaggagaac cctacctaca cggtggacat cttttctgca ggctgcgtct tttactacgt
     2521 aatctctgag ggcagccacc cttttggcaa gtccctgcag cggcaggcca acatcctcct
     2581 gggtgcctgc agccttgact gcttgcaccc agagaagcac gaagacgtca ttgcacgtga
     2641 attgatagag aagatgattg cgatggatcc tcagaaacgc ccctcagcga agcacgtgct
     2701 caaacacccg ttcttctgga gcctagagaa gcagctccag ttcttccagg acgtgagcga
     2761 cagaatagaa aaggaatccc tggatggccc gatcgtgaag cagttagaga gaggcgggag
     2821 agccgtggtg aagatggact ggcgggagaa catcactgtc cccctccaga cagacctgcg
     2881 taaattcagg acctataaag gtggttctgt cagagatctc ctccgagcca tgagaaataa
     2941 gaagcaccac taccgggagc tgcctgcaga ggtgcgggag acgctggggt ccctccccga
     3001 cgacttcgtg tgctacttca cgtctcgctt cccccacctc ctcgcacaca cctaccgggc
     3061 catggagctg tgcagccacg agagactctt ccagccctac tacttccacg agcccccaga
     3121 gccccagccc ccagtgactc cagacgccct ctgagcgagg gcggcccctc tgttctggtg
     3181 gccccagctg tgactgaggg cctggtcaca actagtgttt ctgcagaggc agcaggagcc
     3241 agcccggaat tctgcagata tccatcacac tggcggccgc tcgaccctgt ggaatgtgtg
     3301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3601 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     3661 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3721 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3781 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3841 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3901 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3961 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4021 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4081 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4141 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4201 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     4261 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     4321 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     4381 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     4441 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4501 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     4561 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     4621 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     4681 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     4741 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     4801 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     4861 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     4921 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     4981 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5041 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5101 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5161 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5221 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5281 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5341 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5401 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5461 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5521 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5581 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5641 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5701 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5761 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5821 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5881 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     5941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6961 cctgcagcct gaatatgggc
//
