LOCUS       sequence-90251-e        8120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally FLAG tagged human IRE1a in retroviral pBabe puro 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8120)
  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 8120)
  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_pBabePuro
FEATURES             Location/Qualifiers
     source          1..8120
                     /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(4431..4451)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4436..4765
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4616..4751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4775..5374
                     /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(4775..4794)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5271..5291
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5595..6064
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6194..6211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6365..6953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6445..6464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7113..7904,7905..7973))
                     /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     7736..7755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7974..8078)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(8109..8120,1..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 gcttcctgaa accttgttgt ttgtgtcaac gctggatgga agtttgcatg ctgtcagcaa
     1501 gaggacaggc tcaatcaaat ggactttaaa agaagatcca gtcctgcagg tcccaacaca
     1561 tgtggaagag cctgcctttc tcccagatcc taatgatggc agcctgtata cgcttggaag
     1621 caagaataat gaaggcctga cgaaacttcc ttttaccatc ccagaattgg tgcaggcatc
     1681 cccatgccga agttcagatg gaatcctcta catgggtaaa aagcaggaca tctggtatgt
     1741 tattgacctc ctgaccggag agaagcagca gactttgtca tcggcctttg cagatagtct
     1801 ctgcccatca acctctcttc tgtatcttgg gcgaacagaa tacaccatca ccatgtacga
     1861 caccaaaacc cgagagctcc ggtggaatgc cacctacttt gactatgcgg cctcactgcc
     1921 tgaggacgac gtggactaca agatgtccca ctttgtgtcc aatggtgatg ggctggtggt
     1981 gactgtggac agtgaatctg gggacgtcct gtggatccaa aactacgcct cccctgtggt
     2041 ggccttttat gtctggcagc gggagggtct gaggaaggtg atgcacatca atgtcgctgt
     2101 ggagaccctg cgctatctga ccttcatgtc tggggaggtg gggcgcatca caaagtggaa
     2161 gtacccgttc cccaaggaga cagaggccaa gagcaagctg acgcccactc tgtatgttgg
     2221 gaaatactct accagcctct atgcctctcc ctcaatggta cacgaggggg ttgctgtcgt
     2281 gccccgcggc agcacacttc ctttgctgga agggccccag actgatggcg tcaccatcgg
     2341 ggacaagggg gagtgtgtga tcacgcccag cacggacgtc aagtttgatc ccggactcaa
     2401 aagcaagaac aagctcaact acttgaggaa ttactggctt ctgataggac accatgaaac
     2461 cccactgtct gcgtctacca agatgctgga gagatttccc aacaatctac ccaaacatcg
     2521 ggaaaatgtg attcctgctg attcagagaa aaagagcttt gaggaagtta tcaacctggt
     2581 tgaccagact tcagaaaacg cacctaccac cgtgtctcgg gatgtggagg agaagcccgc
     2641 ccatgcccct gcccggcccg aggcccccgt ggactccatg cttaaggaca tggctaccat
     2701 catcctgagc accttcctgc tgattggctg ggtggccttc atcatcacct atcccctgag
     2761 catgcatcag cagcagcagc tccagcacca gcagttccag aaggaactgg agaagatcca
     2821 gctcctgcag cagcagcagc agcagctgcc cttccaccca cctggagaca cggctcagga
     2881 cggcgagctc ctggacacgt ctggcccgta ctcagagagc tcgggcacca gcagccccag
     2941 cacgtccccc agggcctcca accactcgct ctgctccggc agctctgcct ccaaggctgg
     3001 cagcagcccc tccctggaac aagacgatgg agatgaggaa accagcgtgg tgatagttgg
     3061 gaaaatttcc ttctgtccca aggatgtcct gggccatgga gctgagggca caattgtgta
     3121 ccggggcatg tttgacaacc gcgacgtggc cgtgaagagg atcctccccg agtgttttag
     3181 cttcgcagac cgtgaggtcc agctgttgcg agaatcggat gagcacccga acgtgatccg
     3241 ctacttctgc acggagaagg accggcaatt ccagtacatt gccatcgagc tgtgtgcagc
     3301 caccctgcaa gagtatgtgg agcagaagga ctttgcgcat ctcggcctgg agcccatcac
     3361 cttgctgcag cagaccacct cgggcctggc ccacctccac tccctcaaca tcgttcacag
     3421 agacctaaag ccacacaaca tcctcatatc catgcccaat gcacacggca agatcaaggc
     3481 catgatctcc gactttggcc tctgcaagaa gctggcagtg ggcagacaca gtttcagccg
     3541 ccgatctggg gtgcctggca cagaaggctg gatcgctcca gagatgctga gcgaagactg
     3601 taaggagaac cctacctaca cggtggacat cttttctgca ggctgcgtct tttactacgt
     3661 aatctctgag ggcagccacc cttttggcaa gtccctgcag cggcaggcca acatcctcct
     3721 gggtgcctgc agccttgact gcttgcaccc agagaagcac gaagacgtca ttgcacgtga
     3781 attgatagag aagatgattg cgatggatcc tcagaaacgc ccctcagcga agcacgtgct
     3841 caaacacccg ttcttctgga gcctagagaa gcagctccag ttcttccagg acgtgagcga
     3901 cagaatagaa aaggaatccc tggatggccc gatcgtgaag cagttagaga gaggcgggag
     3961 agccgtggtg aagatggact ggcgggagaa catcactgtc cccctccaga cagacctgcg
     4021 taaattcagg acctataaag gtggttctgt cagagatctc ctccgagcca tgagaaataa
     4081 gaagcaccac taccgggagc tgcctgcaga ggtgcgggag acgctggggt ccctccccga
     4141 cgacttcgtg tgctacttca cgtctcgctt cccccacctc ctcgcacaca cctaccgggc
     4201 catggagctg tgcagccacg agagactctt ccagccctac tacttccacg agcccccaga
     4261 gccccagccc ccagtgactc cagacgccct ctgagcgagg gcggcccctc tgttctggtg
     4321 gccccagctg tgactgaggg cctggtcaca actagtgttt ctgcagaggc agcaggagcc
     4381 agcccggaat tctgcagata tccatcacac tggcggccgc tcgaccctgt ggaatgtgtg
     4441 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4501 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4621 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4681 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4741 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     4801 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4861 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4921 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4981 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5041 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5101 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5161 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5221 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5281 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5341 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     5401 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     5461 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5521 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     5581 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5641 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5701 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     5761 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5821 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5881 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5941 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     6001 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     6061 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     6121 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6181 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6541 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6781 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6961 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7021 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     7081 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7141 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7201 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7261 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7321 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7381 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7441 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7501 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7561 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7621 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7681 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7741 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7801 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7861 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7921 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7981 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8041 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8101 cctgcagcct gaatatgggc
//
