LOCUS       sequence-474222-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses photoactivable fluorescent protein for Mitofusin1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  TITLE     Photoactivable plasmid labeling MFN1 for Mammalian expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6897)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PAmKate-MFN1
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1381..2091)
                     /codon_start=1
                     /product="monomeric photoactivatable red fluorescent 
                     protein (Gunewardene et al., 2011)"
                     /label=PAmKate
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRGD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      2088..2097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2133..2336)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2162..2182)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2337..2640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3732..3779
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3804..3823
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4011..4805)
                     /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(4011..4805)
                     /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(4122..4141)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4732..4751
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4840..5197)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4854..4989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5006..5197
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5154..5174
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5199..5303)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5330..5785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5417..5436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5627..5648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5792..5913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5803..5822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5857..5876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 caaatttctt cccataccat cctctatgtg cttatttaat tcacttttat atatttttaa
       61 tacatctgga ttaggatgaa actctgaaca aaattcatca accaaaacag acagtcgaca
      121 aatttcatct gtcattgcac atgaaacttt gtttgccacc tcctcggtaa cctccttgat
      181 ttttttctta acatccagtg ttaaaaggtt catctggttt cgaataaagt ccagtctatc
      241 aatttggtct tccctctctt ccactgaata atgcctttta tcttcagctg ccaggtttac
      301 tgaatccatt atgtttttca cagtagctag tatctgttta gctctgatag tgtgctgttc
      361 gaactttgtt ttcactgctg actgcgagat acactcctca aagatttgtt caaaattctg
      421 aaattcctgt aatcttgcat gaaatccttc agcaagtgcc acaccacttt ctggcatccc
      481 ctgtgctttt tgctttctag cactaagaac ttcctttgct gaaacaaaga agatacgatt
      541 ctgtgcttct aaagcattta caactttgag ctcctccacc aagaaatgca ggcatctttc
      601 catgtgctgt ctgcgtacgt cttccatata ttctggctct gatgcagagg catcccaacg
      661 attattgaga atgaaaatat taggcttgga aagccgctca ttcaccttgt gaaaaaagtg
      721 tttttccgta ttcattagtg ttgattcaga gtttgcgacc aaaacaaaga catcagcatc
      781 taggcaaaac ttatcaatcc agctatccag ctctgtagtg acatctgtgc ctggactgtc
      841 tactaacacc aggtcatctc tcaagagggc acattttgct tttggccaaa acacacgtac
      901 aagacagcca gctttcaaat ctttgtccat gtgaagggca tgggccagtt gattaactgt
      961 cttcacactc tttttttcat ctgatccttc tgtcataaga taggctttat ctccatcagt
     1021 tccttcaaca cttaggaagc aattggttat atggccaatc ccactaggga gaactttatc
     1081 ccacaacatt gcattgataa cagagctctt cccactgctt gtcctgccaa aaaatgccac
     1141 cttcatgtgt ctccgagata gcacctcacc aatgatggaa agcttgtctt tatatccttg
     1201 catttctacc agatcatctt cagtggctat tcgatcaagt tccggattct tatatgttgc
     1261 ttcaacaaaa tgtgatcctt cagtaacaaa ctccagtaac tggtcaaaga ttgcagtaat
     1321 cgccttctta gccagcacaa agtgcttcag tggagaaaca ggttctgcca tcgaattcga
     1381 attaagcttg tgccccagtt tgctagggag gtcgcagtat ctggccacag ccacctcgtg
     1441 ctgctcgacg taggtctctt tgtcggcctc cttgattctt tccagtctgc ggtccacata
     1501 gtagacgccg ggcatcttga ggttcttagc gggtttcttg gatctgtatg tggtcttcaa
     1561 gttgcagatc aggtggcccc cgcccacgag cttcagggcc atgtcccctc tgccttccag
     1621 gccgccgtca gcggggtaca gcatctcggt gttggcctcc cagccgagtg ttttcttctg
     1681 catcacaggg ccgttggatg ggaagttcac ccctctgatc ttgacgttgt agatgaggca
     1741 gccgtcctgg aggctggtgt cctgggtagc ggtcagcacg cccccgtctt cgtatgtggt
     1801 gactctctcc catgtgaagc cctcagggaa ggactgcttc cagaagtcgg ggatgccctg
     1861 ggtgtggttg atgaaggtct tgctgccgta catgaagctg gtagccagga tgtcgaaggc
     1921 gaaggggaga gggccgccct cgaccacctt gattctcatg gtctgggtgc cctcgtaggg
     1981 cttgccttcg ccctcggatg tgcacttgaa gtggtggttg ttcacggtgc cctccatgta
     2041 cagcttcatg tgcatgttct ccttaatcag ctcttcgccc ttagacacca tggtggcgac
     2101 cggtgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt
     2161 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     2221 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     2281 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     2341 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     2401 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     2461 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     2521 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     2581 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     2641 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     2701 atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3541 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
     3601 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
     3661 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     3721 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     3781 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
     3841 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     3901 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     3961 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     4021 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     4081 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     4141 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     4201 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     4261 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     4321 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     4381 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     4441 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     4501 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     4561 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     4621 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     4681 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     4741 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     4801 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
     4861 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     4921 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     4981 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5041 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5101 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5161 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
     5221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     5341 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5401 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5461 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5521 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5581 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5641 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5701 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5761 aacaaaatat taacgcttac aatttacgcg ttaagataca ttgatgagtt tggacaaacc
     5821 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5881 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     5941 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     6001 ggtatggctg attatgatca gttatctaga tccggtggat ccttaggatt cttcattgct
     6061 tgaaggtaga aactgcttag taaaattctc cagctcattt tcaagttgaa cagctttatt
     6121 tcttaagagc tttgaattgt tttgtatttt ctccaactga tctatttctt tgggtaatct
     6181 agcaatttct tcttccagct gtttttgagt aatatcaact tgttggcaca ggcgagcaaa
     6241 agtggtagct atttgttgtt ttacttggtg actgcagttt gcactcgtgg agctaacaat
     6301 catcctcagt ttttcagttg catagtttac aaactgctgt ttaaaggctc gctccttggc
     6361 atgggtggtc cagctcagtc tttcataaag atacaaagct ccatacatag ttaatgaaac
     6421 agataggagt ttccagccta tagttttcca aatcactcct ccaacaataa tgatgcccat
     6481 agaagttcta gatgtaacgg acgccaatcc tgttactaat gtaatcatga gttcttcctg
     6541 tgatgcatta tctggcgttg ctggagtggt aggagcagtg ggagtagaag ctaaagatct
     6601 agggagctga aagataggct ctgataatcc taggagcacc ctttgagcat ttctagggcc
     6661 caaaaatcga tgtacaaggg aagaccagcc cagggaaaaa cgaaatacaa tatcctcttg
     6721 aaaatctgaa cataacttgt ggtaatttag attataactg agatcaaatt tcttgcaagg
     6781 gatcagtgta tgtagtttat cctgtatacc agctggaagt aatggcttca aattttcaat
     6841 aatttcttgc tgggtctgaa gcactaaggc gtttacttca tcggtgcatc gatcagc
//
