LOCUS       Exported                5668 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses Tom20-mTurquoise2-VASP at the outer mitochondrial membrane
            of mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    OMM PKA Reporter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5668
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        165..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"
     regulatory      1392..1401
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1611..1637
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1638..2354
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1683..1704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1944..1963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2291..2312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2528..2649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2565..2584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2619..2638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2656..3111)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2793..2814)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3005..3024
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3138..3242
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3267..3287)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3311..3521
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3333..3529
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3380..3515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3442..3461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3564..4358
                     /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(3618..3637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4228..4247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4546..4565)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4590..4637
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /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 gtgggccgga acagcgctat tgccgctgga gtgtgcggcg
     1441 ccctgttcat cggctactgc atctacttcg acaggaagag aaggagcgac cccaacttca
     1501 agaacaggct gagggagaga aggaagaagc agaagctggc caaggagaga gccggcctga
     1561 gcaagctgcc cgacctgaag gacgccgagg ccgtgcagaa gttcggtacc tacccatacg
     1621 atgttccaga ttacgctatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1681 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1741 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1801 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
     1861 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1921 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1981 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2041 gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc tatatcaccg
     2101 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     2161 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2221 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     2281 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2341 acgagctgta caagagcgag cacatcgagc ggcgggtgag caacgccggc ggcccccccg
     2401 cctgaggatc ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2461 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2521 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2641 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2701 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2761 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2821 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2881 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2941 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3001 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3061 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3241 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3541 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3721 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4081 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4381 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4441 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4501 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4561 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4621 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4681 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4741 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4801 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5641 tgtggataac cgtattaccg ccatgcat
//
