LOCUS       sequence-202274-        5736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tom20-mTurquoise2-VASP at the outer mitochondrial membrane
            of mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5736)
  AUTHORS   O'Banion CP, Priestman MA, Hughes RM, Herring LE, Capuzzi SJ, 
            Lawrence DS
  TITLE     Design and Profiling of a Subcellular Targeted Optogenetic 
            cAMP-Dependent Protein Kinase.
  JOURNAL   Cell Chem Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011.
  PUBMED    29104065
REFERENCE   2  (bases 1 to 5736)
  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: "Cell Chem 
            Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011."
COMMENT     Sequence Label: OMM PKA Reporter
FEATURES             Location/Qualifiers
     source          1..5736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..1346
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          395..1337
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1294..1314
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1608..1634
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1635..1637,1638..1640,1641..2351)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1680..1701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1941..1960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2288..2309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2524..2645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2561..2580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2615..2634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2652..3107)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2789..2810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3001..3020
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3134..3238
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3240..3597
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3240..3431
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3263..3283)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3448..3583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3632..4426
                     /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             3632..4426
                     /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(3686..3705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4296..4315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4614..4633)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4658..4705
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5034..5622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5523..5542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tgagtaattc atacaaaagg actcgcccct gccttgggga atcccaggga
       61 ccgtcgttaa actcccacta acgtagaacc cagagatcgc tgcgttcccg ccccctcacc
      121 cgcccgctct cgtcatcact gaggtggaga agagcatgcg tgaggctccg gtgcccgtca
      181 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      241 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      301 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      361 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      421 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cgcccctggc
      481 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      541 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggctt gggcgctggg
      601 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      661 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      721 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      781 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      841 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      901 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
      961 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1021 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1081 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1141 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1201 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1261 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1321 tttttcttcc atttcaggtg tcgtgacgct agcgctaccg gactcagatc tcgagctcaa
     1381 gcttcgaatt cgccaccatg atggtgggcc ggaacagcgc tattgccgct ggagtgtgcg
     1441 gcgccctgtt catcggctac tgcatctact tcgacaggaa gagaaggagc gaccccaact
     1501 tcaagaacag gctgagggag agaaggaaga agcagaagct ggccaaggag agagccggcc
     1561 tgagcaagct gcccgacctg aaggacgccg aggccgtgca gaagttctac ccatacgatg
     1621 ttccagatta cgctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1681 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1741 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1801 tgccctggcc caccctcgtg accaccctgt cctggggcgt gcagtgcttc gcccgctacc
     1861 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1921 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1981 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2041 acatcctggg gcacaagctg gagtacaact acatcagcga caacgtctat atcaccgccg
     2101 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2161 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2221 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     2281 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2341 agctgtacaa gagcgagcac atcgagcggc gggtgagcaa cgccggcggc ccccccgcct
     2401 gaggatcccc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2461 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2521 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2581 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2641 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2701 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2761 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2821 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2881 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2941 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3001 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3061 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3121 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3181 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3241 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3301 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3361 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3421 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3481 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3541 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3601 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3661 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3721 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3781 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3841 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3901 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3961 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4021 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4081 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4141 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4201 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4261 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4321 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4381 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4441 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4501 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4561 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4621 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4681 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4741 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4801 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4861 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4921 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4981 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5041 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5101 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5161 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5221 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5281 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5341 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5401 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5461 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5581 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5641 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5701 cctgattctg tggataaccg tattaccgcc atgcat
//
