LOCUS       sequence-431058-        5959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TDP-43 with a C-terminal mCerulean tag for expression in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5959)
  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 5959)
  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 mCerulean
FEATURES             Location/Qualifiers
     source          1..5959
                     /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    1845..1897
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1905..1914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1911..1913,1914..1916,1917..2630)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1956..1977)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2217..2236)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2564..2585
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2844..2863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2881..3336)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3018..3039)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3230..3249
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3363..3467
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3469..3826
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3469..3660
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3492..3512)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3861..4655
                     /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             3861..4655
                     /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(3915..3934)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4525..4544
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4843..4862)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4887..4934
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5263..5851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5752..5771
                     /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 gcatgagcga gtatattcgt gttaccgaag atgagaacga cgagccgatc gagattccga
      661 gcgaagatga cggtacggtc ttgctgagca cggtgaccgc gcagttccca ggcgcctgtg
      721 gcctgcgcta tcgtaatccg gtcagccagt gcatgcgcgg tgtccgtctg gttgagggca
      781 ttctgcacgc tccggacgcg ggctggggca atctggtgta cgtggtcaat tacccgaaag
      841 acaacaagcg caagatggat gaaactgacg ccagctctgc ggttaaagtc aagcgtgcag
      901 tgcagaaaac gagcgacctg atcgtgctgg gtttgccgtg gaaaaccacg gagcaagacc
      961 tgaaagaata ctttagcacc ttcggtgagg tgctgatggt ccaggtaaag aaagatctga
     1021 aaaccggtca cagcaagggt tttggcttcg tccgtttcac cgagtatgaa acccaggtga
     1081 aagttatgtc gcaacgtcac atgatcgacg gtcgttggtg tgactgcaag ctgccgaaca
     1141 gcaagcagag ccaagatgaa ccgctgcgct cccgtaaagt tttcgttggt cgctgcaccg
     1201 aagatatgac ggaagatgag ctgcgtgagt tcttttctca atatggtgac gtgatggatg
     1261 ttttcatccc gaaacctttc cgcgcctttg catttgttac cttcgctgac gatcaaatcg
     1321 cgcagagcct gtgtggtgag gatctgatta tcaaaggtat ttccgttcac atttctaacg
     1381 cggagccaaa gcataacagc aaccgccaac tggaacgttc cggtcgtttc ggtggtaatc
     1441 cgggtggttt cggcaaccag ggtggctttg gcaattcgcg tggcggtggt gccggcttgg
     1501 gcaataacca aggttccaat atgggtggcg gtatgaactt tggtgccttc tccatcaatc
     1561 cggcaatgat ggcggctgca caagcggcgc tgcagtctag ctggggtatg atgggcatgc
     1621 tggcgagcca gcagaaccag tcaggcccga gcggcaataa tcagaatcaa ggtaacatgc
     1681 agcgtgaacc gaatcaagcg tttggtagcg gtaacaatag ctacagcggt agcaacagcg
     1741 gtgcggcaat cggctggggt agcgcaagca atgcgggcag cggtagcggt ttcaacggcg
     1801 gctttggtag cagcatggat agcaagagct ccggttgggg catgctcgag ctcaagcttc
     1861 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggtgagca
     1921 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1981 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2041 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2101 ccctgacctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     2161 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2221 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2281 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2341 acaacgccat cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     2401 ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2461 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2521 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2581 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     2641 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2701 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2881 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2941 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3001 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3061 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3121 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3181 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3241 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3301 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3481 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3841 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3901 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3961 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4021 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4081 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4141 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4201 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4261 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4321 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4381 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4441 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4501 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4561 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4621 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4681 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4741 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4801 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4861 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4921 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4981 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5041 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5101 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5161 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5881 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5941 ggataaccgt attaccgcc
//
