LOCUS       sequence-366422-        7279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-level transient expression of full-length HsIRE1 with a 
            C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7279)
  AUTHORS   Belyy V, Zuazo-Gaztelu I, Alamban A, Ashkenazi A, Walter P
  TITLE     Endoplasmic reticulum stress activates human IRE1alpha through 
            reversible assembly of inactive dimers into small oligomers.
  JOURNAL   Elife. 2022 Jun 22;11:e74342. doi: 10.7554/eLife.74342.
  PUBMED    35730415
REFERENCE   2  (bases 1 to 7279)
  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"; journalName: "Elife."; 
            date: "2022-06-22"; volume: "11:e74342. doi"; pages: " 
            10.7554/eLife.74342"
COMMENT     Sequence Label: pPW3758 : CMVd3-HsIRE1-HaloTag
FEATURES             Location/Qualifiers
     source          1..7279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..136
                     /label=CMVd3 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields strongly reduced expression 
                     relative to the full-length promoter"
     CDS             3148..4038
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    4170..4291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4207..4226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4261..4280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4298..4753)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4435..4456)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4647..4666
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4780..4884
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4909..4929)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4953..5163
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4975..5171
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5022..5157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5206..6000
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5206..6000
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5260..5279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5870..5889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6188..6207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6232..6279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6608..7196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7097..7116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gatgggcggt aggcgtgtac ggtgggaggt
      121 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
      181 atgccggccc ggcggctgct gctgctgctg acgctgctgc tgcccggcct cgggattttt
      241 ggaagtacca gcacagtgac gcttcctgaa accttgttgt ttgtgtcaac gctggatgga
      301 agtttgcatg ctgtcagcaa gaggacaggc tcaatcaaat ggactttaaa agaagatcca
      361 gtcctgcagg tcccaacaca tgtggaagag cctgcctttc tcccagatcc taatgatggc
      421 agcctgtata cgcttggaag caagaataat gaaggcctga cgaaacttcc ttttaccatc
      481 ccagaattgg tgcaggcatc cccatgccga agttcagatg gaatcctcta catgggtaaa
      541 aagcaggaca tctggtatgt tattgacctc ctgaccggag agaagcagca gactttgtca
      601 tcggcctttg cagatagtct ctgcccatca acctctcttc tgtatcttgg gcgaacagaa
      661 tacaccatca ccatgtacga caccaaaacc cgagagctcc ggtggaatgc cacctacttt
      721 gactatgcgg cctcactgcc tgaggacgac gtggactaca agatgtccca ctttgtgtcc
      781 aatggtgatg ggctggtggt gactgtggac agtgaatctg gggacgtcct gtggatccaa
      841 aactacgcct cccctgtggt ggccttttat gtctggcagc gggagggtct gaggaaggtg
      901 atgcacatca atgtcgctgt ggagaccctg cgctatctga ccttcatgtc tggggaggtg
      961 gggcgcatca caaagtggaa gtacccgttc cccaaggaga cagaggccaa gagcaagctg
     1021 acgcccactc tgtatgttgg gaaatactct accagcctct atgcctctcc ctcaatggta
     1081 cacgaggggg ttgctgtcgt gccccgcggc agcacacttc ctttgctgga agggccccag
     1141 actgatggcg tcaccatcgg ggacaagggg gagtgtgtga tcacgcccag cacggacgtc
     1201 aagtttgatc ccggactcaa aagcaagaac aagctcaact acttgaggaa ttactggctt
     1261 ctgataggac accatgaaac cccactgtct gcgtctacca agatgctgga gagatttccc
     1321 aacaatctac ccaaacatcg ggaaaatgtg attcctgctg attcagagaa aaagagcttt
     1381 gaggaagtta tcaacctggt tgaccagact tcagaaaacg cacctaccac cgtgtctcgg
     1441 gatgtggagg agaagcccgc ccatgcccct gcccggcccg aggcccccgt ggactccatg
     1501 cttaaggaca tggctaccat catcctgagc accttcctgc tgattggctg ggtggccttc
     1561 atcatcacct atcccctgag catgcatcag cagcagcagc tccagcacca gcagttccag
     1621 aaggaactgg agaagatcca gctcctgcag cagcagcagc agcagctgcc cttccaccca
     1681 cctggagaca cggctcagga cggcgagctc ctggacacgt ctggcccgta ctcagagagc
     1741 tcgggcacca gcagccccag cacgtccccc agggcctcca accactcgct ctgctccggc
     1801 agctctgcct ccaaggctgg cagcagcccc tccctggaac aagacgatgg agatgaggaa
     1861 accagcgtgg tgatagttgg gaaaatttcc ttctgtccca aggatgtcct gggccatgga
     1921 gctgagggca caattgtgta ccggggcatg tttgacaacc gcgacgtggc cgtgaagagg
     1981 atcctccccg agtgttttag cttcgcagac cgtgaggtcc agctgttgcg agaatcggat
     2041 gagcacccga acgtgatccg ctacttctgc acggagaagg accggcaatt ccagtacatt
     2101 gccatcgagc tgtgtgcagc caccctgcaa gagtatgtgg agcagaagga ctttgcgcat
     2161 ctcggcctgg agcccatcac cttgctgcag cagaccacct cgggcctggc ccacctccac
     2221 tccctcaaca tcgttcacag agacctaaag ccacacaaca tcctcatatc catgcccaat
     2281 gcacacggca agatcaaggc catgatctcc gactttggcc tctgcaagaa gctggcagtg
     2341 ggcagacaca gtttcagccg ccgatctggg gtgcctggca cagaaggctg gatcgctcca
     2401 gagatgctga gcgaagactg taaggagaac cctacctaca cggtggacat cttttctgca
     2461 ggctgcgtct tttactacgt aatctctgag ggcagccacc cttttggcaa gtccctgcag
     2521 cggcaggcca acatcctcct gggtgcctgc agccttgact gcttgcaccc agagaagcac
     2581 gaagacgtca ttgcacgtga attgatagag aagatgattg cgatggatcc tcagaaacgc
     2641 ccctcagcga agcacgtgct caaacacccg ttcttctgga gcctagagaa gcagctccag
     2701 ttcttccagg acgtgagcga cagaatagaa aaggaatccc tggatggccc gatcgtgaag
     2761 cagttagaga gaggcgggag agccgtggtg aagatggact ggcgggagaa catcactgtc
     2821 cccctccaga cagacctgcg taaattcagg acctataaag gtggttctgt cagagatctc
     2881 ctccgagcca tgagaaataa gaagcaccac taccgggagc tgcctgcaga ggtgcgggag
     2941 acgctggggt ccctccccga cgacttcgtg tgctacttca cgtctcgctt cccccacctc
     3001 ctcgcacaca cctaccgggc catggagctg tgcagccacg agagactctt ccagccctac
     3061 tacttccacg agcccccaga gccccagccc ccagtgactc cagacgccct cggtggaggc
     3121 ggtagcggag gaggtggctc tgtcgacatg gcagaaatcg gtactggctt tccattcgac
     3181 ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat
     3241 ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc
     3301 atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa
     3361 tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc
     3421 atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg
     3481 ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc
     3541 atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc
     3601 ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt
     3661 acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag
     3721 ccgttcctga atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc
     3781 gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag
     3841 tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa
     3901 gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg
     3961 aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg
     4021 acgctcgaga tttccggcta agatccaccg gatctagata actgatcata atcagccata
     4081 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4141 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4201 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4261 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     4321 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4381 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4441 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4501 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4561 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4621 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4681 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4741 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4861 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4921 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5101 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5161 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5221 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5281 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5341 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5401 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5461 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5521 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5581 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5641 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5701 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5761 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5821 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5881 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5941 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6001 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6061 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6241 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6421 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7261 ttcctgcgtt atcccctga
//
