LOCUS       sequence-459165-        6077 bp    DNA     circular SYN 15-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 6077)
  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 6077)
  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: 
            "10.1016/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..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..18)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(172..760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(252..271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(931..1722,1723..1791))
                     /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     1554..1573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1792..1896)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1964..1982
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2020..2042)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2142..2161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2355..2377
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2369..2386
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2370..2386
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2413..2716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2717..2920
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2871..2891
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2917..2941
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3058..3154
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(3386..3388,3389..3391,3392..3850)
                     /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(3431..3452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3692..3711)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    5602..5736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5626..5645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5680..5699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5847..5863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5847..5863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5860..5882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5871..5887
                     /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(5895..5901,5902..5919,5920..5925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5940..5961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
       61 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1921 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     1981 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2041 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2101 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     2161 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2221 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     2281 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     2341 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc
     2401 atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2461 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2521 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2581 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2641 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2701 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2761 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2821 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2881 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     2941 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     3001 cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag ttaactggta
     3061 agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga
     3121 actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct
     3181 ctaaaagctg cggaattgta cccgcgggcc caccatgccg gcccggcggc tgctgctgct
     3241 gctgacgctg ctgctgcccg gcctcggggc atcaatgcag aagctgatct cagaggagga
     3301 cctgcttatg gccatggagg cccgaattcg gtcgaccgag atctctcgag gtaccggagg
     3361 tggcgggagc ggaggtggcg ggagtatggt gagcaagggc gaggagctgt tcaccggggt
     3421 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3481 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     3541 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     3601 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3661 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3721 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3781 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3841 tctcaccgcc atttttggaa gtaccagcac agtgacgctt cagacttcag aaaacgcacc
     3901 taccaccgtg tctcgggatg tggaggagaa gcccgcccat gcccctgccc ggcccgaggc
     3961 ccccgtggac tccatgctta aggacatggc taccatcatc ctgagcacct tcctgctgat
     4021 tggctgggtg gccttcatca tcacctatcc cctgagcatg catcagcagc agcagctcca
     4081 gcaccagcag ttccagaagg aactggagaa gatccagctc ctgcagcagc agcagcagca
     4141 gctgcccttc cacccacctg gagacacggc tcaggacggc gagctcctgg acacgtctgg
     4201 cccgtactca gagagctcgg gcaccagcag ccccagcacg tcccccaggg cctccaacca
     4261 ctcgctctgc tccggcagct ctgcctccaa ggctggcagc agcccctccc tggaacaaga
     4321 cgatggagat gaggaaacca gcgtggtgat agttgggaaa atttccttct gtcccaagga
     4381 tgtcctgggc catggagctg agggcacaat tgtgtaccgg ggcatgtttg acaaccgcga
     4441 cgtggccgtg aagaggatcc tccccgagtg ttttagcttc gcagaccgtg aggtccagct
     4501 gttgcgagaa tcggatgagc acccgaacgt gatccgctac ttctgcacgg agaaggaccg
     4561 gcaattccag tacattgcca tcgagctgtg tgcagccacc ctgcaagagt atgtggagca
     4621 gaaggacttt gcgcatctcg gcctggagcc catcaccttg ctgcagcaga ccacctcggg
     4681 cctggcccac ctccactccc tcaacatcgt tcacagagac ctaaagccac acaacatcct
     4741 catatccatg cccaatgcac acggcaagat caaggccatg atctccgact ttggcctctg
     4801 caagaagctg gcagtgggca gacacagttt cagccgccga tctggggtgc ctggcacaga
     4861 aggctggatc gctccagaga tgctgagcga agactgtaag gagaacccta cctacacggt
     4921 ggacatcttt tctgcaggct gcgtctttta ctacgtaatc tctgagggca gccacccttt
     4981 tggcaagtcc ctgcagcggc aggccaacat cctcctgggt gcctgcagcc ttgactgctt
     5041 gcacccagag aagcacgaag acgtcattgc acgtgaattg atagagaaga tgattgcgat
     5101 ggatcctcag aaacgcccct cagcgaagca cgtgctcaaa cacccgttct tctggagcct
     5161 agagaagcag ctccagttct tccaggacgt gagcgacaga atagaaaagg aatccctgga
     5221 tggcccgatc gtgaagcagt tagagagagg cgggagagcc gtggtgaaga tggactggcg
     5281 ggagaacatc actgtccccc tccagacaga cctgcgtaaa ttcaggacct ataaaggtgg
     5341 ttctgtcaga gatctcctcc gagccatgag aaataagaag caccactacc gggagctgcc
     5401 tgcagaggtg cgggagacgc tggggtccct ccccgacgac ttcgtgtgct acttcacgtc
     5461 tcgcttcccc cacctcctcg cacacaccta ccgggccatg gagctgtgca gccacgagag
     5521 actcttccag ccctactact tccacgagcc cccagagccc cagcccccag tgactccaga
     5581 cgccttgtga gcggccgcgg ggatccagac atgataagat acattgatga gtttggacaa
     5641 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     5701 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     5761 atgtttcagg ttcaggggga ggtgtgggag gttttttcgg atcctctaga gtcgatctgc
     5821 aggcatgcta gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5881 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5941 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6001 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6061 gggcgctctt ccgcttc
//
