LOCUS       sequence-251151-        7868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER-stress sensor - XBP1 splicing fluorescent reporter with 
            mNeonGreen (retroviral vector backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7868)
  AUTHORS   Nougarede A, Tesniere C, Ylanko J, Rimokh R, Gillet G, Andrews DW
  TITLE     Improved IRE1 and PERK Pathway Sensors for Multiplex Endoplasmic 
            Reticulum Stress Assay Reveal Stress Response to Nuclear Dyes Used 
            for Image Segmentation.
  JOURNAL   Assay Drug Dev Technol. 2018 Aug 8. doi: 10.1089/adt.2018.862.
  PUBMED    30088945
REFERENCE   2  (bases 1 to 7868)
  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: "Assay Drug 
            Dev Technol. 2018 Aug 8. doi: 10.1089/adt.2018.862."
COMMENT     Sequence Label: pLHCX-XBP1 mNeonGreen NLS
FEATURES             Location/Qualifiers
     source          1..7868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     639..659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     685..709
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             800..826
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             800..826
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1087..1791
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      1339..1348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1813..1839
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     LTR             1894..2487
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2590..2612)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2698..2838
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2753..2772
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2853..2870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3024..3612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3104..3123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3783..4574,4575..4643))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4406..4425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4644..4748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4816..4834
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5406..5581
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5644..5843
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6044..6460
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6390..6412
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6428..6444
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6516..7535
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6516..7535
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
ORIGIN
        1 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc catatcataa
       61 tatgtacatt tatattggct catgtccaac attaccgcca tgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccc
      781 caagcttgaa ttcaccatgt acccctacga cgtgcccgac tacgccggcc ttgtagttga
      841 gaaccaggag ttaagacagc gcttggggat ggatgccctg gttgctgaag aggaggcgga
      901 agccaagggg aatgaagtga ggccagtggc cgggtctgct gagtccgcag cactcagact
      961 acgtgcacct ctgcagcagg tgcaggccca gttgtcaccc ctccagaaca tctccccttg
     1021 gattctggcg gtattgactc ttcagattca gagtctgata tcctgttggg cattctaata
     1081 ggatccgtga gcaagggcga ggaggataac atggcctctc tcccagcgac acatgagtta
     1141 cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg caccggcaat
     1201 ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct ccagttctcc
     1261 ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc ctaccctgac
     1321 gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt ccatcgcaca
     1381 atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta cgagggaagc
     1441 cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg
     1501 accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc caacgacaaa
     1561 accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg ctaccggagc
     1621 actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct gaagaaccag
     1681 ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct caacttcaag
     1741 gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa ggtcgacggt
     1801 ggatcagggg gacccgccgc caagcgcgtg aagctggact aatctagaat cgataaaata
     1861 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     1921 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     1981 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     2041 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     2101 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     2161 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     2221 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     2281 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     2341 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     2401 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     2461 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     2521 cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt tcggtgatga
     2581 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2641 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     2701 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     2761 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2821 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2881 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3421 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     3481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4081 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4381 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     4441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4741 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4801 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attcatacca
     4861 gatcaccgaa aactgtcctc caaatgtgtc cccctcacac tcccaaattc gcgggcttct
     4921 gcctcttaga ccactctacc ctattcccca cactcaccgg agccaaagcc gcggcccttc
     4981 cgtttctttg cttttgaaag accccacccg taggtggcaa gctagcttaa gtaacgccac
     5041 tttgcaaggc atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa
     5101 caaagaaaca gctgaatacc aaacaggata tctgtggtaa gcggttcctg ccccggctca
     5161 gggccaagaa cagatgagac agctgagtga tgggccaaac aggatatctg tggtaagcag
     5221 ttcctgcccc ggctcggggc caagaacaga tggtccccag atgcggtcca gccctcagca
     5281 gtttctagtg aatcatcaga tgtttccagg gtgccccaag gacctgaaaa tgaccctgta
     5341 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tccgctctcc
     5401 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcttccg atagactgcg
     5461 tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggtct
     5521 cgctgttcct tgggagggtc tcctctgagt gattgactac ccacgacggg ggtctttcat
     5581 ttgggggctc gtccgggatt tggagacccc tgcccaggga ccaccgaccc accaccggga
     5641 ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt
     5701 tatgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg
     5761 aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg
     5821 ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata
     5881 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     5941 ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg
     6001 ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac
     6061 tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc
     6121 ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa
     6181 cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa
     6241 ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga
     6301 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
     6361 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
     6421 atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattccga tctgatagct
     6481 tgccacaacc cgtaccaaag atggatagat ccggaaagcc tgaactcacc gcgacgtctg
     6541 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     6601 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     6661 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     6721 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     6781 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     6841 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     6901 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     6961 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     7021 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     7081 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     7141 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     7201 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     7261 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     7321 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     7381 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     7441 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     7501 gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg aacaagagct
     7561 gatttcgaga acgcctcagc cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa
     7621 cggccttacg cttggtggca cagttctcgt ccacagttcg ctaagctcgc tcggctgggt
     7681 cgcgggaggg ccggtcgcag tgattcaggc ccttctggat tgtgttggtc cccagggcac
     7741 gattgtcatg cccacgcact cgggtgatct gactgatccc gcagattgga gatcgccgcc
     7801 cgtgcctgcc gattgggtgc agatctctag agctcgctga tccggccatt agccatatta
     7861 ttcattgg
//
