LOCUS       sequence-458834-        7482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses control plasmid for ER-mitochondrial linkers, tagged with 
            mRFP, from a lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7482)
  AUTHORS   Dematteis G, Tapella L, Casali C, Talmon M, Tonelli E, Reano S, 
            Ariotti A, Pessolano E, Malecka J, Chrostek G, Kulkoviene G, 
            Umbrasas D, Distasi C, Grilli M, Ladds G, Filigheddu N, Fresu LG, 
            Mikoshiba K, Matute C, Ramos-Gonzalez P, Jekabsone A, Cali T, Brini 
            M, Biggiogera M, Cavaliere F, Miggiano R, Genazzani AA, Lim D
  TITLE     ER-mitochondria distance is a critical parameter for efficient 
            mitochondrial Ca(2+) uptake and oxidative metabolism.
  JOURNAL   Commun Biol. 2024 Oct 10;7(1):1294. doi: 10.1038/s42003-024-06933-9.
  PUBMED    39390051
REFERENCE   2  (bases 1 to 7482)
  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: "2024-10-10"; volume: "7(1):1294. doi"; pages: " 
            10.1038/s42003-024-06933-9"
COMMENT     Sequence Label: pLV-ER-mRFP
FEATURES             Location/Qualifiers
     source          1..7482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     566..590
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             665..1339
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             665..1339
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(795..813)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1063..1082
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1257..1280
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    1447..2035
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1500..1520)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1918..1929)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2122..2355
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2427..2548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2464..2483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2518..2537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2588..2723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2743..2762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2744..2762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2772..2789)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2772..2788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2781..2803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2930..3385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3017..3036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3227..3248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3411..3515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3516..3584,3585..4376)
                     /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     complement(3734..3753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4547..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5036..5055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5289..5306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5423..5444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5459..5464,5465..5482,5483..5489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5497..5513
                     /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)."
     primer_bind     5502..5524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5521..5537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5521..5537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5556..5576
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5558..5576
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5604..5830
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5831..6011
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6058..6183
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6676..6909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7094..7138
                     /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    join(7405..7482,1..39)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 taaagaatta caaaaacaaa ttacaaaatt caaaatttta tcgataagct tgggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      601 gccatccacg ctgttttgac ctccatagaa gacaccgact ctagaggatc caccggtcgc
      661 caccatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga
      721 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      781 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
      841 catcctgtcc cctcagttcc agtacggctc caaggcctac gtgaagcacc ccgccgacat
      901 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
      961 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1021 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1081 gaccatgggc tgggaggcct ccaccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1141 cgagatcaag atgaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac
     1201 cacctacatg gccaagaagc ccgtgcagct gcccggcgcc tacaagaccg acatcaagct
     1261 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg
     1321 ccgccactcc accggcgcct ccggactcag atctcgagct caagcttcga attcccgggt
     1381 catggtttat attggtatcg ctattttttt gtttgttggc ctttttatga aataggtacc
     1441 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1501 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1561 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1621 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1681 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1741 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1801 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1861 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     1921 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1981 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2041 attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     2101 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     2161 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2341 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     2401 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     2461 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2521 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     2581 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2641 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2701 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     2761 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2821 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2881 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     2941 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3001 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3061 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3121 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3181 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3241 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3301 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3361 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3481 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3541 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3601 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3661 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3721 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3781 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3841 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3901 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3961 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4021 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4081 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4141 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4201 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4261 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4321 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4381 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4441 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4501 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4561 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4621 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5161 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5221 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5281 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5341 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5401 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5461 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5521 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     5581 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
     5641 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
     5701 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     5761 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     5821 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5881 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5941 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6001 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     6061 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     6121 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     6181 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     6241 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     6301 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     6361 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     6421 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     6481 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     6541 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     6601 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     6661 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     6721 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     6781 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     6841 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     6901 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     6961 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     7021 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     7081 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     7141 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     7201 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     7261 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     7321 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     7381 cggatctcga cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc
     7441 aggggaaaga atagtagaca taatagcaac agacatacaa ac
//
