LOCUS       sequence-382390-        6870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic tool.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6870)
  AUTHORS   Guillen-Samander A, Leonzino M, Hanna MG, Tang N, Shen H, De Camilli
            P
  TITLE     VPS13D bridges the ER to mitochondria and peroxisomes via Miro.
  JOURNAL   J Cell Biol. 2021 May 3;220(5). pii: 212021. doi: 
            10.1083/jcb.202010004.
  PUBMED    33891013
REFERENCE   2  (bases 1 to 6870)
  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: "2021-05-3"; volume: "220(5). pii: 212021. doi"; 
            pages: " 10.1083/jcb.202010004"
COMMENT     Sequence Label: mCh-Miro1(∆TM)-SspB
FEATURES             Location/Qualifiers
     source          1..6870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             612..1319
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(757..775)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1025..1044
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3183..3512
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     polyA_signal    3658..3779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3695..3714)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3749..3768
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3786..4241)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3923..3944)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4135..4154
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4268..4372
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4374..4731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4374..4565
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4397..4417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4582..4717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4766..5560
                     /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             4766..5560
                     /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(4820..4839)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5430..5449
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5748..5767)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5792..5839
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6168..6756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6657..6676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
      661 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      721 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      781 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      841 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      901 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      961 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1021 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1081 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1141 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1201 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1261 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga
     1321 gatctcgagc tcaagcttcg aatttgggat ctatgaagaa agacgtgcgg atcctgctgg
     1381 tgggagaacc tagagttggg aagacatcac tgattatgtc tctggtcagt gaagaatttc
     1441 cagaagaggt tcctccccgg gcagaagaaa tcaccattcc agctgatgtc accccagaga
     1501 gagttccaac acacattgta gattactcag aagcagaaca gagtgatgaa caacttcatc
     1561 aagaaatatc tcaggctaat gtcatctgta tagtgtatgc cgttaacaac aagcattcta
     1621 ttgataaggt aacaagtcga tggattcctc tcataaatga aagaacagac aaagacagca
     1681 ggctgccttt aatattggtt gggaacaaat ctgatctggt ggaatatagt agtatggaga
     1741 ccatccttcc tattatgaac cagtatacag aaatagaaac ctgtgtggag tgttcagcga
     1801 aaaacctgaa gaacatatca gagctctttt attacgcaca gaaagctgtt cttcatccta
     1861 cagggcccct gtactgccca gaggagaagg agatgaaacc agcttgtata aaagccctta
     1921 ctcgtatatt taaaatatct gatcaagata atgatggtac tctcaatgat gctgaactca
     1981 acttctttca gaggatttgt ttcaacactc cattagctcc tcaagctctg gaggatgtca
     2041 agaatgtagt cagaaaacat ataagtgatg gtgtggctga cagtgggttg accctgaaag
     2101 gttttctctt tttacacaca ctttttatcc agagagggag acacgaaact acttggactg
     2161 tgcttcgacg atttggttat gatgatgacc tggatttgac acctgaatat ttgttccccc
     2221 tgctgaaaat acctcctgat tgcactactg aattaaatca tcatgcatat ttatttctcc
     2281 aaagcacctt tgacaagcat gatttggata gagactgtgc tttgtcacct gatgagctta
     2341 aagatttatt taaagttttc ccttacatac cttgggggcc agatgtgaat aacacagttt
     2401 gtaccaatga aagaggctgg ataacctacc agggattcct ttcccagtgg acgctcacga
     2461 cttatttaga tgtacagcgg tgcctggaat atttgggcta tctaggctat tcaatattga
     2521 ctgagcaaga gtctcaagct tcagctgtta cagtgacaag agataaaaag atagacctgc
     2581 agaaaaaaca aactcaaaga aatgtgttca gatgtaatgt aattggagtg aaaaactgtg
     2641 ggaaaagtgg agttcttcag gctcttcttg gaagaaactt aatgaggcag aagaaaattc
     2701 gtgaagatca taaatcctac tatgcgatta acactgttta tgtatatgga caagagaaat
     2761 acttgttgtt gcatgatatc tcagaatcgg aatttctaac tgaagctgaa atcatttgtg
     2821 atgttgtatg cctggtatat gatgtcagca atcccaaatc ctttgaatac tgtgccagga
     2881 tttttaagca acactttatg gacagcagaa taccttgctt aatcgtagct gcaaagtcag
     2941 acctgcatga agttaaacaa gaatacagta tttcacctac tgatttctgc aggaaacaca
     3001 aaatgcctcc accacaagcc ttcacttgca atactgctga tgcccccagt aaggatatct
     3061 ttgttaaatt gacaacaatg gccatgtatc cgcacgtgac acaagctgac ctcaagagct
     3121 ctcgagctca agcttcgaac gaattcggca ttgatctgag cggcctgacc ctgcaggaat
     3181 tcagctcccc gaaacgccct aagctgctgc gtgaatatta cgattggctg gttgataaca
     3241 gctttacccc atatctggtg gtggatgcca catacctggg cgtgaacgtg cccgtggagt
     3301 atgtgaaaga cggtcagatc gtgctgaatc tgtctgcaag tgcgaccggc aacctgcaac
     3361 tgacaaatga ttttatccag ttcaacgccc gctttaaggg cgtgtctcgt gaactgtata
     3421 tcccgatggg tgccgctctg gccatttacg ctcgcgagaa cggcgatggt gtgatgttcg
     3481 aaccagaaga aatctatgac gagctgaata ttggttaacg cgggcccggg atccaccgga
     3541 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3601 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3661 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3721 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3781 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3841 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3901 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3961 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4021 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4081 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4141 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4201 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4261 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4321 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4381 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4741 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4801 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4861 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4921 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4981 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5041 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5101 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5161 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5221 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5281 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5341 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5401 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5461 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5521 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5581 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5641 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5701 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5761 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5821 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5881 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5941 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6001 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6061 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6121 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6181 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6241 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6301 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6361 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6421 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6481 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6541 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6601 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6661 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6721 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6781 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6841 tctgtggata accgtattac cgccatgcat
//
