LOCUS       sequence-352602-        7439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DM Firefly luciferase reporter fused to a prolactin signal sequence 
            for ER targeting and KDEL for ER retention and fused to eGFP at the 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  AUTHORS   Cortes Sanchon A, Santhosh Kumar H, Mantovani M, Osinnii I, Mateos 
            JM, Kaech A, Shcherbakov D, Akbergenov R, Bottger EC
  TITLE     ER-misfolded proteins become sequestered with mitochondria and 
            impair mitochondrial function.
  JOURNAL   Commun Biol. 2021 Dec 2;4(1):1350. doi: 10.1038/s42003-021-02873-w.
  PUBMED    34857875
REFERENCE   2  (bases 1 to 7439)
  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: "Commun 
            Biol."; date: "2021-12-2"; volume: "4(1):1350. doi"; pages: " 
            10.1038/s42003-021-02873-w"
COMMENT     Sequence Label: pPLN-hFluc-DM-eGFP-KDEL
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..1077
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1056..1064
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    1101..1149
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1305..1322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1476..2064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1556..1575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2264..3055,3056..3124))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3229..3277
                     /note="synthetic polyadenylation signal"
     misc_feature    3291..3382
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3331..3350
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3533..3912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3913..4116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4067..4087
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4113..4137
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4309..5955
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDQDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFQVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             join(5992..5994,5995..5997,5998..6708)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5992..5994,5995..5997,5998..6708)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6037..6058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6298..6317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6645..6666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6776..6897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6787..6806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6841..6860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        join(7091..7439,1..9)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7091..7282
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7114..7134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7299..7434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7361..7380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
       61 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
      121 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
      181 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
      241 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
      301 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
      361 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
      421 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
      481 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
      541 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
      601 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
      661 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
      721 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
      781 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
      841 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
      901 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
      961 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     1021 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttaaact
     1081 ctagaaccgg tcatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     1141 ttttgtgtgt tcgaactaga tgctgtcgac cgatgccctt gagagccttc aacccagtca
     1201 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     1261 tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg
     1321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2221 acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag tgcttgatca
     2281 gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc tgactccccg
     2341 tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca gcaatgatgc
     2401 cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca gcagggaggg
     2461 ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg agctgctgtc
     2521 gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg gccattgcta
     2581 ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct ggttcccagc
     2641 ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc tccttagggc
     2701 ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta atggcagcac
     2761 tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc ggcgagtact
     2821 caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta
     2881 tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc gggaatcgtt
     2941 cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg atatagccca
     3001 ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa
     3061 aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc
     3121 tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag tacgtctctc
     3181 aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa taaaatatct
     3241 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     3301 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     3361 gcaggtgcca gaacatttct ctggcctaac tggcctcaat attggccatt agccatatta
     3421 ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     3481 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttggcattga
     3541 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     3601 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     3661 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     3721 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     3781 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3841 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     3901 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     3961 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4021 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     4081 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact
     4141 agaagcttgg caacagtacc gtgatacaac cacactcgag ccatgaacat caaaggatcg
     4201 ccatggaaag ggtccctcct gctgctgctg gtgtcaaacc tgctcctgtg ccagagcgtg
     4261 gccccctgga attcagcttg gcaatccggt actgttggta aagccaccga agatgccaaa
     4321 aacattaaga agggcccagc gccattctac ccactcgaag acgggaccgc cggcgagcag
     4381 ctgcacaaag ccatgaagcg ctacgccctg gtgcccggca ccatcgcctt taccgacgca
     4441 catatcgagg tggacattac ctacgccgag tacttcgaga tgagcgttcg gctggcagaa
     4501 gctatgaagc gctatgggct gaatacaaac catcggatcg tggtgtgcag cgagaatagc
     4561 ttgcagttct tcatgcccgt gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct
     4621 aacgacatct acaacgagcg cgagctgctg aacagcatgg gcatcagcca gcccaccgtc
     4681 gtattcgtga gcaagaaagg gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc
     4741 atacaaaaga tcatcatcat ggatagcaag accgactacc agggcttcca aagcatgtac
     4801 accttcgtga cttcccattt gccacccggc ttcaacgagt acgacttcgt gcccgagagc
     4861 ttcgaccagg acaaaaccat cgccctgatc atgaacagta gtggcagtac cggattgccc
     4921 aagggcgtag ccctaccgca ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc
     4981 atcttcggca accagatcat ccccgacacc gctatcctca gcgtggtgcc atttcaccac
     5041 ggcttcggca tgttcaccac gctgggctac ttgatctgcg gctttcaagt cgtgctcatg
     5101 taccgcttcg aggaggagct attcttgcgc agcttgcaag actataagat tcaatctgcc
     5161 ctgctggtgc ccacactatt tagcttcttc gctaagagca ctctcatcga caagtacgac
     5221 ctaagcaact tgcacgagat cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag
     5281 gccgtggcca aacgcttcca cctaccaggc atccgccagg gctacggcct gacagaaaca
     5341 accagcgcca ttctgatcac ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg
     5401 gtgcccttct tcgaggctaa ggtggtggac ttggacaccg gtaagacact gggtgtgaac
     5461 cagcgcggcg agctgtgcgt ccgtggcccc atgatcatga gcggctacgt taacaacccc
     5521 gaggctacaa acgctctcat cgacaaggac ggctggctgc acagcggcga catcgcctac
     5581 tgggacgagg acgagcactt cttcatcgtg gaccggctga agagcctgat caaatacaag
     5641 ggctaccagg tagccccagc cgaactggag agcatcctgc tgcaacaccc caacatcttc
     5701 gacgccgggg tcgccggcct gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc
     5761 gtgctggaac acggtaaaac catgaccgag aaggagatcg tggactatgt ggccagccag
     5821 gttacaaccg ccaagaagct gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga
     5881 ctgaccggca agttggacgc ccgcaagatc cgcgagattc tcattaaggc caagaagggc
     5941 ggcaagatcg ccgtgatccc cgggatcggt ggtggttctg gaggcggttc tatggtgagc
     6001 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     6061 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     6121 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     6181 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     6241 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     6301 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     6361 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     6421 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     6481 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     6541 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     6601 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     6661 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagaa aaagaccaaa
     6721 gatgaactct aataattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     6781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     6901 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     6961 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtttgc gtattgggcg
     7021 ctcttccgct gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     7081 tagctacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     7141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7201 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     7261 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     7321 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     7381 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggc
//
