LOCUS       sequence-471594-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SNAP-tag fused to wild-type TDP-43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  TITLE     Shelkovnikova lab plasmids 2025
REFERENCE   2  (bases 1 to 6549)
  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"
COMMENT     Sequence Label: SNAP-tag TDP-43
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..81
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             144..689
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     regulatory      2031..2040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2037..2039,2040..2042,2043..2756)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2037..2039,2040..2042,2043..2753)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2082..2103)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2343..2362)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2690..2711
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2879..3000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2916..2935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2970..2989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3007..3462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3356..3375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3489..3593
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3595..3952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3595..3786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3618..3638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3803..3938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3987..4781
                     /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             3987..4781
                     /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(4041..4060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4651..4670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4969..4988)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5013..5060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5389..5977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5878..5897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6152..6455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6456..6549,1..110)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
       61 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      121 accgtcagat ccgctagccc accatggaca aagactgcga aatgaagcgc accaccctgg
      181 atagccctct gggcaagctg gaactgtctg ggtgcgaaca gggcctgcac cgtatcatct
      241 tcctgggcaa aggaacatct gccgccgacg ccgtggaagt gcctgcccca gccgccgtgc
      301 tgggcggacc agagccactg atgcaggcca ccgcctggct caacgcctac tttcaccagc
      361 ctgaggccat cgaggagttc cctgtgccag ccctgcacca cccagtgttc cagcaggaga
      421 gctttacccg ccaggtgctg tggaaactgc tgaaagtggt gaagttcgga gaggtcatca
      481 gctacagcca cctggccgcc ctggccggca atcccgccgc caccgccgcc gtgaaaaccg
      541 ccctgagcgg aaatcccgtg cccattctga tcccctgcca ccgggtggtg cagggcgacc
      601 tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga gtggctgctg gcccacgagg
      661 gccacagact gggcaagcct gggctgggtc ctgcaggcgg atccggtacc gaattcgcgg
      721 ccgcagctcg agctatgtct gaatatattc gggtaaccga agatgagaac gatgagccca
      781 ttgaaatacc atcggaagac gatgggacgg tgctgctctc cacggttaca gcccagtttc
      841 caggggcgtg tgggcttcgc tacaggaatc cagtgtctca gtgtatgaga ggtgtccggc
      901 tggtagaagg aattctgcat gccccagatg ctggctgggg aaatctggtg tatgttgtca
      961 actatccaaa agataacaaa agaaaaatgg atgagacaga tgcttcatca gcagtgaaag
     1021 tgaaaagagc agtccagaaa acatccgatt taatagtgtt gggtctccca tggaaaacaa
     1081 ccgaacagga cctgaaagag tattttagta cctttggaga agttcttatg gtgcaggtca
     1141 agaaagatct taagactggt cattcaaagg ggtttggctt tgttcgtttc acggaatatg
     1201 aaacacaagt gaaagtaatg tcacagcgac atatgataga tggacgatgg tgtgactgca
     1261 aacttcctaa ttctaagcaa agccaagatg agcctttgag aagcagaaaa gtgtttgtgg
     1321 ggcgctgtac agaggacatg actgaggatg agctgcggga gttcttctct cagtacgggg
     1381 atgtgatgga tgtcttcatc cccaagccat tcagggcctt tgcctttgtt acatttgcag
     1441 atgatcagat tgcgcagtct ctttgtggag aggacttgat cattaaagga atcagcgttc
     1501 atatatccaa tgccgaacct aagcacaata gcaatagaca gttagaaaga agtggaagat
     1561 ttggtggtaa tccaggtggc tttgggaatc agggtggatt tggtaatagc agagggggtg
     1621 gagctggttt gggaaacaat caaggtagta atatgggtgg tgggatgaac tttggtgcgt
     1681 tcagcattaa tccagccatg atggctgccg cccaggcagc actacagagc agttggggta
     1741 tgatgggcat gttagccagc cagcagaacc agtcaggccc atcgggtaat aaccaaaacc
     1801 aaggcaacat gcagagggag ccaaaccagg ccttcggttc tggaaataac tcttatagtg
     1861 gctctaattc tggtgcagca attggttggg gatcagcatc caatgcaggg tcgggcagtg
     1921 gttttaatgg aggctttggc tcaagcatgg attctaagtc ttctggctgg ggaatgtaga
     1981 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg
     2041 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2101 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2161 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2221 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2281 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2341 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2401 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2461 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2521 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2581 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2641 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2701 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     2761 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2821 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3001 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3061 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3481 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3541 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3601 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3781 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3901 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3961 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4021 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4081 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4141 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4201 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4261 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4321 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4381 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4441 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4501 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4561 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4621 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4681 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4741 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4801 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4861 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4921 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4981 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5041 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5101 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5161 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5221 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6061 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     6121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     6481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     6541 atgggagtt
//
