LOCUS       sequence-440863-        4289 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  Expresses the VN fragment of BiFC-IRE1-dLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4289)
  AUTHORS   Cho H, Stanzione F, Oak A, Kim GH, Yerneni S, Qi L, Sum AK, Chan C
  TITLE     Intrinsic Structural Features of the Human IRE1alpha Transmembrane 
            Domain Sense Membrane Lipid Saturation.
  JOURNAL   Cell Rep. 2019 Apr 2;27(1):307-320.e5. doi: 
            10.1016/j.celrep.2019.03.017.
  PUBMED    30943411
REFERENCE   2  (bases 1 to 4289)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2019.03"; journalName: "Cell Rep"; date: 
            "2019-04-2- 2"; volume: "27"; issue: "1"; pages: "307-320.e5"
COMMENT     Sequence Label: pBiFC-SS-VN155(I152L)-IRE1-dLD
FEATURES             Location/Qualifiers
     source          1..4289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(121..123,124..126,127..585)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(166..187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(427..446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    600..734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(624..643)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     678..697
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(845..861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(845..861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(858..880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    869..885
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(893..899,900..917,918..923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    938..959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1076..1093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1247..1835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1327..1346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2006..2797,2798..2866))
                     /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     2629..2648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2867..2971)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3039..3057
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3095..3117)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3217..3236
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3430..3452
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3444..3461
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3445..3461
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3488..3791
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3792..3995
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3946..3966
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3992..4016
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4133..4229
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 atggcatcaa tgcagaagct gatctcagag gaggacctgc ttatggccat ggaggcccga
       61 attcggtcga ccgagatctc tcgaggtacc ggaggtggcg ggagcggagg tggcgggagt
      121 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      181 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      241 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      301 ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      361 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      421 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      481 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      541 aagctggagt acaactacaa cagccacaac gtctatctca ccgcctgagc ggccgcgggg
      601 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      661 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      721 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      781 tgtgggaggt tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat
      841 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
      901 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
      961 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1021 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1681 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3001 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3061 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3181 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     3241 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3301 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     3361 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     3421 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc
     3481 tgcaggtcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     3541 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     3601 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     3661 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     3721 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     3781 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     3841 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     3901 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     3961 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     4021 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     4081 ggactctaga ggatccggta ctagaggaac tgaaaaacca gaaagttaac tggtaagttt
     4141 agtctttttg tcttttattt caggtcccgg atccggtggt ggtgcaaatc aaagaactgc
     4201 tcctcagtgg atgttgcctt tacttctagg cctgtacgga agtgttactt ctgctctaaa
     4261 agctgcggaa ttgtacccgc gggcccacc
//
