LOCUS       sequence-417776-        5487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mScarlet-I-PDCD10 targeted to the mitochondria in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5487)
  AUTHORS   Ball NJ, Ghimire S, Follain G, Pajari AO, Wurzinger D, Vaitkeviciute
            M, Cowell AR, Berki B, Ivaska J, Paatero I, Goult BT, Jacquemet G
  TITLE     TLNRD1 is a CCM complex component and regulates endothelial barrier 
            integrity.
  JOURNAL   J Cell Biol. 2024 Sep 2;223(9):e202310030. doi: 
            10.1083/jcb.202310030. Epub 2024 Jul 16.
  PUBMED    39013281
REFERENCE   2  (bases 1 to 5487)
  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: "J Cell 
            Biol."; date: "2024-09-2"; volume: "223(9):e202310030. doi"; pages: 
            " 10.1083/jcb.202310030. Epub 2024 Jul 16"
COMMENT     Sequence Label: mito-PDCD10-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..5487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     526..545
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(634..991)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      648..783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        800..991
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     948..968
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(993..1097)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1124..1579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1211..1230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1421..1442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1586..1707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1597..1616)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1651..1670
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2528..3223)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2803..2822)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3072..3090
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3220..3229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3257..3331)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     complement(3350..3369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3351..3369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3414..3617)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3443..3463)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3618..3921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5013..5060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5085..5104
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(5292..5487,1..599))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(join(5292..5487,1..599))
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5403..5422)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
       61 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      121 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      181 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
      241 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
      301 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
      361 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
      421 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
      481 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
      541 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
      601 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
      661 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
      721 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
      781 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      841 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      901 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      961 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
     1021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1561 atattaacgc ttacaattta cgccttaaga tacattgatg agtttggaca aaccacaact
     1621 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1681 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1741 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1801 gctgattatg atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc
     1861 agaattccta tgccactgtc ttaaacgttt gcaagatgag gttggtttgg tgaataaggc
     1921 gattcgcgct tacgaagacg ttaattgctt ttccgtcttt gaagtaagtc ttcaaagtat
     1981 cactgaaaga ctttgagtac ttcacgaact ccttcttttg gtgttcgagt gcccttctat
     2041 tttgatactg gtacttcttg aatacgttgt tcacggtatc caacagttcc tttatggcgg
     2101 aggcaatgtc tttgatagtt tgcagaaacc gtaccctatc gttgatctcg tccgggattt
     2161 ttgagagaat ttgcttcagc gcccgagctt tttcgttgag atcttggaac tcaggctcag
     2221 gtctctcgat catatactcc tcaacgtcat cggcggccat ccgcagcaga gactcggtga
     2281 aatttacctc cacagatttt ttttcaagaa ttttcattat gatgtcttga gtcaggccgg
     2341 ggttttcctt ttctgccttg atgaacgccg cccgcaatgt ctgtgctgct gaaagattca
     2401 cccgttcgag ctcgttaaaa actggataca ttacggcgta caaaggcatg gataccatag
     2461 acgtggtttc ggcttcattc ttcatttctt ccattgtcat cctcatagct cgagatctga
     2521 gtccggactt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gagcgttcgt
     2581 actgttccac cacggtgtag tcctcgttgt gggaggtgat gtccaacttg cggtcgacgt
     2641 tgtaggcgcc gggcatctgc acgggcttct tggccttgta ggtggtcttg aagtccgcca
     2701 ggtagcggcc gccgtccttc aggcgcaggg ccatcttaat gtcgcccttc agcacgccgt
     2761 cctcggggta caaccgctcg gtggacgctt cccagcccat tgtcttcttc tgcattacgg
     2821 ggccgtcagg agggaagttg gtgccgcgga gcttcacctt gtagatcagg gtgccgtcct
     2881 ccagggaggt gtcctgggtc acggtcacgg cgccgccgtc ctcgaagttc atcacgcgct
     2941 cccacttgaa gccctcgggg aaggactgct tatagtagtc ggggatgtcg gcggggtgct
     3001 tgatgaaggc cctggagccg tacatgaact gaggggacag gatgtcccag gagaagggca
     3061 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     3121 cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg tgcaccttga
     3181 accgcatgaa ctccttgatc actgcctcgc ccttgctcac catggtggcg accggtggat
     3241 cccccaacga atggatcttg gcgcgcggca ctgggagccg ccgggccgag cctgtcaagc
     3301 cccgcagcag cagcggcgtc aggacggaca tgctagccag cttgggtctc cctatagtga
     3361 gtcgtattaa tttcgataag ccagtaagca gtgggttctc tagttagcca gagagctctg
     3421 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     3481 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     3541 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     3601 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     3661 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     3721 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     3781 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     3841 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     3901 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     3961 attgattact attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     4861 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     4921 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     4981 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     5041 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     5101 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     5161 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     5221 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     5281 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     5341 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     5401 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     5461 cagtcgatga atccagaaaa gcggcca
//
