LOCUS       sequence-458399-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stabilizes the ER-mitochondrial distance at 15 nm; tagged with mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  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 6592)
  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: pcDNA3.1-15nmEML-mRFP
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1198..1869
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     primer_bind     complement(1325..1343)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2183..2200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2189..2413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2459..2887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2546..2565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2756..2777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2896..2916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2901..3230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3081..3216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3143..3162
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3297..4091
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3351..3370)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3961..3980
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4265..4386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4302..4321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4356..4375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4435..4451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4435..4451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4448..4470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4459..4475
                     /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(join(4483..4489,4490..4507,4508..4513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4666..4683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4837..5425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4917..4936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5596..6387,6388..6456))
                     /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     6219..6238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6457..6561)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atggctattc agctgcggtc tctgttccct ctggctctgc ctggactgct ggctctgctg
      961 gggtggtggt ggttcttctc ccgaaaaaag gaccccaccc ggtccgccaa catggagaag
     1021 gagagaaagc ggagagagga ggatgagcag aggcgccgga aggaggagga ggagagaagg
     1081 atgaagctgg agatggaggc caagcggaag caggaggagg aggagcggaa gaagagagag
     1141 gacgatgaga agcggatcca ggccgagaga aacagcgacc cacctgtggc cacactggcc
     1201 agctccgagg atgtgatcaa ggagttcatg cggtttaaag tgagaatgga gggctccgtg
     1261 aatggccacg agttcgagat cgagggagag ggagagggca gaccttacga gggcacccag
     1321 acagccaagc tgaaggtgac caagggagga ccactgccct tcgcctggga catcctgtct
     1381 cctcagtttc agtacggcag caaggcctat gtgaagcacc ctgccgacat cccagattat
     1441 ctgaagctga gcttcccaga gggctttaag tgggagaggg tgatgaactt tgaggatggc
     1501 ggcgtggtga ccgtgacaca ggactctagc ctgcaggatg gcgagttcat ctacaaggtg
     1561 aagctgcgcg gcacaaattt tccctccgac ggccctgtga tgcagaagaa gaccatgggc
     1621 tgggaggcct ctacagagag gatgtatccc gaggatggcg ccctgaaggg cgagatcaag
     1681 atgcgcctga agctgaagga cggcggccac tacgatgccg aggtgaagac cacatatatg
     1741 gccaagaagc cagtgcagct gccaggagcc tacaagaccg acatcaagct ggatatcaca
     1801 tctcacaacg aggactacac catcgtggag cagtatgaga gggccgaggg ccgccactct
     1861 accggagcca gcggcctgag gtcccgcgcc caggcctcca attctatgga gaaggagcgg
     1921 aagcgccggg aggaggatga gcagagaagg cgcaaagaag aggaggagcg gagaatgaaa
     1981 ctggagatgg aagccaagag aaaacaggaa gaggaggaga ggaagaagcg cgaggacgat
     2041 gagaagagga tccaggcaga gagccgcgcc caggccagca attcccgagt gatggtctac
     2101 attggaatcg ctatctttct gttcgtggga ctgtttatga agtgactcga gtctagaggg
     2161 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2221 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2281 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2341 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2401 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2581 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2821 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4081 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4141 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4201 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4261 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4381 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4441 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
