LOCUS       sequence-431056-        5981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TDP-43 with a C-terminal mRuby tag and Lysine 84 in the NLS mutated 
            to a TAG stop codon for amber codon suppression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5981)
  AUTHORS   Shadish JA, Lee JC
  TITLE     Genetically encoded lysine photocage for spatiotemporal control of 
            TDP-43 nuclear import.
  JOURNAL   Biophys Chem. 2024 Jan 23;307:107191. doi: 
            10.1016/j.bpc.2024.107191.
  PUBMED    38290242
REFERENCE   2  (bases 1 to 5981)
  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: "Biophys 
            Chem."; date: "2024-01-23"; volume: "307:107191. doi"; pages: " 
            10.1016/j.bpc.2024.107191"
COMMENT     Sequence Label: TDP-43 mRuby K84ONBK
FEATURES             Location/Qualifiers
     source          1..5981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    597..677
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1969..2649
                     /codon_start=1
                     /product="monomeric variant of the red fluorescent protein 
                     eqFP611 (Kredel et al., 2009)"
                     /label=mRuby
                     /translation="MNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKV
                     IEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVITV
                     MQDTSLEDGCLVYHAQVRGVNFPSNGAVMQKKTKGWEPNTEMMYPADGGLRGYTHMALK
                     VDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVAKFA
                     GLGGG"
     polyA_signal    2775..2896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2812..2831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2866..2885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2903..3358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3040..3061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3252..3271
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3385..3489
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3491..3848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3491..3682
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3514..3534)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3699..3834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3883..4677
                     /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             3883..4677
                     /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(3937..3956)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4547..4566
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4865..4884)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4909..4956
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5285..5873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5774..5793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
      661 gcccgggatc caccggtcgc caccatgagc gagtatattc gtgttaccga agatgagaac
      721 gacgagccga tcgagattcc gagcgaagat gacggtacgg tcttgctgag cacggtgacc
      781 gcgcagttcc caggcgcctg tggcctgcgc tatcgtaatc cggtcagcca gtgcatgcgc
      841 ggtgtccgtc tggttgaggg cattctgcac gctccggacg cgggctgggg caatctggtg
      901 tacgtggtca attacccgaa agacaacaag cgctagatgg atgaaactga cgccagctct
      961 gcggttaaag tcaagcgtgc agtgcagaaa acgagcgacc tgatcgtgct gggtttgccg
     1021 tggaaaacca cggagcaaga cctgaaagaa tactttagca ccttcggtga ggtgctgatg
     1081 gtccaggtaa agaaagatct gaaaaccggt cacagcaagg gttttggctt cgtccgtttc
     1141 accgagtatg aaacccaggt gaaagttatg tcgcaacgtc acatgatcga cggtcgttgg
     1201 tgtgactgca agctgccgaa cagcaagcag agccaagatg aaccgctgcg ctcccgtaaa
     1261 gttttcgttg gtcgctgcac cgaagatatg acggaagatg agctgcgtga gttcttttct
     1321 caatatggtg acgtgatgga tgttttcatc ccgaaacctt tccgcgcctt tgcatttgtt
     1381 accttcgctg acgatcaaat cgcgcagagc ctgtgtggtg aggatctgat tatcaaaggt
     1441 atttccgttc acatttctaa cgcggagcca aagcataaca gcaaccgcca actggaacgt
     1501 tccggtcgtt tcggtggtaa tccgggtggt ttcggcaacc agggtggctt tggcaattcg
     1561 cgtggcggtg gtgccggctt gggcaataac caaggttcca atatgggtgg cggtatgaac
     1621 tttggtgcct tctccatcaa tccggcaatg atggcggctg cacaagcggc gctgcagtct
     1681 agctggggta tgatgggcat gctggcgagc cagcagaacc agtcaggccc gagcggcaat
     1741 aatcagaatc aaggtaacat gcagcgtgaa ccgaatcaag cgtttggtag cggtaacaat
     1801 agctacagcg gtagcaacag cggtgcggca atcggctggg gtagcgcaag caatgcgggc
     1861 agcggtagcg gtttcaacgg cggctttggt agcagcatgg atagcaagag ctccggttgg
     1921 ggcatgctcg agggttctgc tggttctgct gctggttctg gtgaattcat gaacagcctg
     1981 atcaaagaaa acatgcggat gaaggtggtg ctggaaggca gcgtgaacgg ccaccagttc
     2041 aagtgcaccg gcgagggcga gggcaacccc tacatgggca cccagaccat gcggatcaaa
     2101 gtgatcgagg gcggacctct gcccttcgcc ttcgacatcc tggccacatc cttcatgtac
     2161 ggcagccgga ccttcatcaa gtaccccaag ggcatccccg atttcttcaa gcagagcttc
     2221 cccgagggct tcacctggga gagagtgacc agatacgagg acggcggcgt gatcaccgtg
     2281 atgcaggaca ccagcctgga agatggctgc ctggtgtacc atgcccaggt caggggcgtg
     2341 aattttccca gcaacggcgc cgtgatgcag aagaaaacca agggctggga gcccaacacc
     2401 gagatgatgt accccgctga cggcggactg agaggctaca cccacatggc cctgaaggtg
     2461 gacggcggag ggcacctgag ctgcagcttc gtgaccacct accgatccaa gaaaaccgtg
     2521 ggcaacatca agatgcccgg catccacgcc gtggaccacc ggctggaaag gctggaagag
     2581 tccgacaacg agatgttcgt ggtgcagcgg gagcacgccg tggccaagtt cgccggcctg
     2641 ggcggagggt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     2701 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2761 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2821 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2881 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2941 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3001 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3061 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3121 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3181 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3241 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3301 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3361 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3481 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3541 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3601 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3661 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3721 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3781 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3841 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3901 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3961 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4021 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4081 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4141 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4201 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4261 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4321 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4381 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4441 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4501 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4561 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4621 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4681 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4741 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4801 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4861 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4921 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4981 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5041 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5101 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5941 ctgcgttatc ccctgattct gtggataacc gtattaccgc c
//
