LOCUS       sequence-222082-        7973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER-stress sensor - ATF4 translation at ORF3 fluorescent reporter 
            with mScarlet-I (retroviral vector backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7973)
  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 7973)
  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-ATF4 mScarlet NLS
FEATURES             Location/Qualifiers
     source          1..7973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        483..686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     683..707
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1154..1180
                     /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             1154..1180
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1916..1942
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     LTR             1997..2590
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2693..2715)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2801..2941
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2856..2875
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2956..2973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3127..3715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3207..3226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3886..4677,4678..4746))
                     /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     4509..4528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4747..4851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4919..4937
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5509..5684
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5747..5946
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6147..6563
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6493..6515
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6531..6547
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6619..7638
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6619..7638
                     /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 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcca tatcataata
       61 tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
      721 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cgcggcccca
      781 agcttgaatt ctctactttg cccgcccaca gatgtagttt tctctgcgcg tgtgcgtttt
      841 ccctcctccc cgccctcagg gtccacggcc accatggcgt attaggggca gcagtgcctg
      901 cggcagcatt ggcctttgca gcggcggcag cagcaccagg ctctgcagcg gcaaccccca
      961 gcggcttaag ccatggcgct tctcacggca ttcagcagca gcgttgctgt aaccgacaaa
     1021 gacaccttcg aattaagcac attcctcgat tccagcaaag caccgcaaca tgaccgaaat
     1081 gagcttcctg agcagcgagg tgttggtggg ggacttgatg tcccccttcg accagtcggg
     1141 tttgggggct gaatacccct acgacgtgcc cgactacgcc ggtggatcag ggggaggatc
     1201 cgtctccaaa ggggaagcgg tcattaaaga gttcatgagg ttcaaagttc atatggaggg
     1261 cagtatgaac ggccatgaat ttgaaatcga gggcgagggg gagggacgcc cttacgaagg
     1321 gacacaaaca gccaaactga aggttacgaa aggtgggcca cttcccttta gttgggatat
     1381 actgtcacct caatttatgt acggcagccg agccttcatt aaacatcccg ctgacattcc
     1441 ggattactat aagcagtcct ttccagaggg gtttaaatgg gagcgggtta tgaatttcga
     1501 ggatggcggg gccgtaactg tcacgcagga taccagtctg gaagatggta ccctgattta
     1561 taaggttaag ctcagaggaa cgaacttccc gccagatggt cccgtaatgc aaaaaaagac
     1621 gatgggttgg gaggcatcaa ctgaaagatt gtatcctgag gatggtgtcc ttaagggtga
     1681 tattaaaatg gctctgaggc tgaaggatgg cggaaggtat ttggcagatt tcaaaactac
     1741 atataaggcc aagaagcctg ttcagatgcc aggtgcctat aatgtggaca gaaaacttga
     1801 cattacatca cacaacgagg attataccgt cgtggagcaa tatgagagaa gtgagggccg
     1861 acatagtacc ggcggcatgg atgaactcta taaggtcgac ggtggatcag ggggacccgc
     1921 cgccaagcgc gtgaagctgg actaatctag aatcgataaa ataaaagatt ttatttagtc
     1981 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     2041 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     2101 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     2161 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     2221 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     2281 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     2341 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     2401 ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
     2461 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     2521 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     2581 gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca
     2641 ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2701 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2761 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2821 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     2881 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2941 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3001 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3061 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3541 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4201 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4501 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4861 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4921 taggcgtatc acgaggccct ttcgtcttca agaattcata ccagatcacc gaaaactgtc
     4981 ctccaaatgt gtccccctca cactcccaaa ttcgcgggct tctgcctctt agaccactct
     5041 accctattcc ccacactcac cggagccaaa gccgcggccc ttccgtttct ttgcttttga
     5101 aagaccccac ccgtaggtgg caagctagct taagtaacgc cactttgcaa ggcatggaaa
     5161 aatacataac tgagaataga aaagttcaga tcaaggtcag gaacaaagaa acagctgaat
     5221 accaaacagg atatctgtgg taagcggttc ctgccccggc tcagggccaa gaacagatga
     5281 gacagctgag tgatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcgg
     5341 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gtgaatcatc
     5401 agatgtttcc agggtgcccc aaggacctga aaatgaccct gtaccttatt tgaactaacc
     5461 aatcagttcg cttctcgctt ctgttcgcgc gcttccgctc tccgagctca ataaaagagc
     5521 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
     5581 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
     5641 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
     5701 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
     5761 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
     5821 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
     5881 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     5941 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     6001 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     6061 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     6121 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     6181 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     6241 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     6301 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     6361 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     6421 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     6481 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     6541 ccctcactcc ttctctaggc gccggaattc cgatctgata gcttgccaca acccgtacca
     6601 aagatggata gatccggaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     6661 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     6721 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     6781 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     6841 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     6901 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     6961 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     7021 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     7081 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     7141 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     7201 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     7261 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     7321 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     7381 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     7441 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     7501 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     7561 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     7621 ccgagggcaa aggaatagag tagatgccga ccgaacaaga gctgatttcg agaacgcctc
     7681 agccagcaac tcgcgcgagc ctagcaaggc aaatgcgaga gaacggcctt acgcttggtg
     7741 gcacagttct cgtccacagt tcgctaagct cgctcggctg ggtcgcggga gggccggtcg
     7801 cagtgattca ggcccttctg gattgtgttg gtccccaggg cacgattgtc atgcccacgc
     7861 actcgggtga tctgactgat cccgcagatt ggagatcgcc gcccgtgcct gccgattggg
     7921 tgcagatctc tagagctcgc tgatccggcc attagccata ttattcattg gtt
//
