LOCUS Exported 9066 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ER-stress sensor - XBP1 splicing fluorescent reporter with mNeonGreen. ACCESSION . VERSION . KEYWORDS pLVX-XBP1 mNeonGreen NLS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9066) 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 9066) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9066 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 12..315 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 316..519 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 470..490 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 516..540 /label=LNCX /note="Human CMV promoter, forward primer" CDS 654..680 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 941..1645 /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 1193..1202 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1667..1693 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" promoter 1713..2212 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(1748..1771) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 2145..2164 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 2233..2832 /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(2233..2252) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2729..2749 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2846..3434 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2899..2919) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3317..3328) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3641..4274 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(4402..4418) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4402..4418) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4415..4437) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4426..4442 /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(4450..4480) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4495..4516 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4633..4650) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4804..5392) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4884..4903) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5563..6423) /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 6186..6205 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6424..6528) /gene="bla" /label=AmpR promoter polyA_signal 6576..6710 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6613..6632) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6667..6686 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 6878..7511 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 7558..7683 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8180..8413 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8598..8642 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 8905..9020 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" ORIGIN 1 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 61 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 121 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 181 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 241 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 301 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 361 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 421 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 481 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc 541 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctactagagg 601 atcgctagcg ctaccggact cagatctcga gctcaagctt cgaattcacc atgtacccct 661 acgacgtgcc cgactacgcc ggccttgtag ttgagaacca ggagttaaga cagcgcttgg 721 ggatggatgc cctggttgct gaagaggagg cggaagccaa ggggaatgaa gtgaggccag 781 tggccgggtc tgctgagtcc gcagcactca gactacgtgc acctctgcag caggtgcagg 841 cccagttgtc acccctccag aacatctccc cttggattct ggcggtattg actcttcaga 901 ttcagagtct gatatcctgt tgggcattct aataggatcc gtgagcaagg gcgaggagga 961 taacatggcc tctctcccag cgacacatga gttacacatc tttggctcca tcaacggtgt 1021 ggactttgac atggtgggtc agggcaccgg caatccaaat gatggttatg aggagttaaa 1081 cctgaagtcc accaagggtg acctccagtt ctccccctgg attctggtcc ctcatatcgg 1141 gtatggcttc catcagtacc tgccctaccc tgacgggatg tcgcctttcc aggccgccat 1201 ggtagatggc tccggatacc aagtccatcg cacaatgcag tttgaagatg gtgcctccct 1261 tactgttaac taccgctaca cctacgaggg aagccacatc aaaggagagg cccaggtgaa 1321 ggggactggt ttccctgctg acggtcctgt gatgaccaac tcgctgaccg ctgcggactg 1381 gtgcaggtcg aagaagactt accccaacga caaaaccatc atcagtacct ttaagtggag 1441 ttacaccact ggaaatggca agcgctaccg gagcactgcg cggaccacct acacctttgc 1501 caagccaatg gcggctaact atctgaagaa ccagccgatg tacgtgttcc gtaagacgga 1561 gctcaagcac tccaagaccg agctcaactt caaggagtgg caaaaggcct ttaccgatgt 1621 gatgggcatg gacgagctgt acaaggtcga cggtggatca gggggacccg ccgccaagcg 1681 cgtgaagctg gactaatcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca 1741 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg 1801 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc 1861 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc 1921 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag 1981 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc 2041 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg 2101 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca 2161 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag 2221 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 2281 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 2341 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 2401 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 2461 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 2521 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 2581 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 2641 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 2701 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 2761 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 2821 cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt 2881 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 2941 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 3001 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 3061 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 3121 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 3181 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 3241 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 3301 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 3361 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 3421 ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc 3481 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag 3541 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca 3601 gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc 3661 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat 3721 tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac 3781 aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc 3841 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg 3901 aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc 3961 aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg 4021 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct 4081 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 4141 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 4201 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 4261 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 4321 gaaatgaata tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag 4381 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6541 aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg 6601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 6661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa 6721 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt 6781 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa 6841 attgtatttg ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa 6901 agaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag 6961 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag 7021 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg 7081 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg 7141 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag 7201 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt 7261 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt 7321 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 7381 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 7441 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 7501 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc 7561 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg 7621 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc 7681 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg 7741 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc 7801 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta 7861 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc 7921 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata 7981 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca 8041 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag 8101 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag 8161 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca 8221 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag 8281 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca 8341 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg 8401 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc 8461 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga 8521 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat 8581 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt 8641 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag 8701 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta 8761 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca 8821 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag 8881 tgaacggatc tcgacggtat cgcctttaaa agaaaagggg ggattggggg gtacagtgca 8941 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa 9001 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttatcga 9061 taagct //