LOCUS       sequence-333973-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for transcription of mutant TDP-43-M337V mRNA for injection 
            into zebrafish embryos for BiFC assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  AUTHORS   Don EK, Maschirow A, Radford RAW, Scherer NM, Vidal-Itriago A, Hogan
            A, Maurel C, Formella I, Stoddart JJ, Hall TE, Lee A, Shi B, Cole 
            NJ, Laird AS, Badrock AP, Chung RS, Morsch M
  TITLE     In vivo Validation of Bimolecular Fluorescence Complementation 
            (BiFC) to Investigate Aggregate Formation in Amyotrophic Lateral 
            Sclerosis (ALS).
  JOURNAL   Mol Neurobiol. 2021 May;58(5):2061-2074. doi: 
            10.1007/s12035-020-02238-0. Epub 2021 Jan 7.
  PUBMED    33415684
REFERENCE   2  (bases 1 to 5316)
  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"; doi: 
            "10.1007/s12035-020-02238-0"; journalName: "Mol Neurobiol"; date: 
            "2021-05"; volume: "58"; issue: "5"; pages: "2061-2074"
COMMENT     Sequence Label: pCS2plus-TDP-43 M337V
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    781..915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(818..837)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     872..891
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(2230..2248)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2231..2248)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2283..3267)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3338..3793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3425..3444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3635..3656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3819..3923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3924..3992,3993..4784)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4142..4161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4955..5316,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgctcccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg ggtaccgggc ccaatgcatt ggcgccgcgg ccgcgaatta
      721 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
      781 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      901 tatcatgtct ggatctacgt aatacgactc actatagttc tagactacat tccccagcca
      961 gaagacttag aatccatgct tgagccaaag cctccattaa aaccactgcc cgaccctgca
     1021 ttggatgctg atccccaacc aattgctgca ccagaattag agccactata agagttattt
     1081 ccagaaccga aggcctggtt tggctccctc tgcatgttgc cttggttttg gttattaccc
     1141 gatgggcctg actggttctg ctggctggct aacatgccca ccatacccca actgctctgt
     1201 agtgctgcct gggcggcagc catcatggct ggattaatgc tgaacgcacc aaagttcatc
     1261 ccaccaccca tattactacc ttgattgttt cccaaaccag ctccaccccc tctgctatta
     1321 ccaaatccac cctgattccc aaagccacct ggattaccac caaatcttcc acttctttct
     1381 aactgtctat tgctattgtg cttaggttcg gcattggata tatgaacgct gattccttta
     1441 atgatcaagt cctctccaca aagagactgc gcaatctgat catctgcaaa tgtaacaaag
     1501 gcaaaggccc tgaatggctt ggggatgaag acatccatca catccccgta ctgagagaag
     1561 aactcccgca gctcatcctc agtcatgtcc tctgtacagc gccccacaaa cacttttctg
     1621 cttctcaaag gctcatcttg gctttgctta gaattaggaa gtttgcagtc acaccatcgt
     1681 ccatctatca tatgtcgctg tgacattact ttcacttgtg tttcatattc cgtaaaacga
     1741 acaaagccaa acccctttga atgaccagtc ttaagatctt tcttgacctg caccataaga
     1801 acttctccaa aggtactaaa atactctttc aggtcctgtt cggttgtttt ccatgggaga
     1861 cccaacacta ttaaatcgga tgttttctgg actgctcttt tcactttcac tgctgatgaa
     1921 gcatctgtct catccatttt tcttttgtta tcttttggat agttgacaac atacaccaga
     1981 tttccccagc cagcatctgg ggcatgcaga attccttcta ccagccggac acctctcata
     2041 cactgagaca ctggattcct gtagcgaagc ccacacgccc ctggaaactg ggctgtaacc
     2101 gtggagagca gcaccgtccc atcgtcttcc gatggtattt caatgggctc atcgttctca
     2161 tcttcggtta cccgaatata ttcagacatg gtggcggcgg atcctgcaaa aagaacaagt
     2221 agcttgtatt ctatagtgtc acctaaatca agcttgctcc gacgtcccca ggcagaatgg
     2281 cggttcccta aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt
     2341 tacgtcaatg gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgt
     2401 acttggcagc catcgcgggc catttaccgc cattgacgtc aatgggagta ctgccaatgt
     2461 accctggcgt acttccaata gtaatgtact tgccaagtta ctattaatag atattgatgt
     2521 actgccaagt gggccattta ccgtcattga cgtcaatagg gggcgtgaga acggatatga
     2581 atgggcaatg agccatccca ttgacgtcaa tggtgggtgg tcctattgac gtcaatgggc
     2641 attgagccag gcgggccatt taccgtaatt gacgtcaatg ggggaggcgc catatacgtc
     2701 aataggaccg cccatatgac gtcaatagga aagaccatat atagagacca ttgacgtcaa
     2761 tgggggagtg gctatgggcg gtattaggaa gccccatata tggtatatgg gaccgcccat
     2821 tgggaggggc tatctacgtc aataggaaaa cccatatatg gaatactata tggcataggg
     2881 ccaatacata gtattgaacc tggccaatag ccatattggc atagggccat attggatatt
     2941 gcctatatat tgatcctggc atatagccaa tatggccgcc attattggca ccatgccaat
     3001 tcaatatggc ggacttggca cagggccaag acccctcctc aaacttggca ccgtgccaag
     3061 tagacccctc cccagttggc acagtgccag gtccgccata ttgaattggc atggtgccaa
     3121 gtccgccata ttgaattggc tggggtccag atccaccata ttgaattggc acgagtccat
     3181 atacgccata ttgaattggc acaggtccag atccaccata ttgaattggc atgagtccat
     3241 atacgccata ttgaattggc ggtcgactgg cgtaatagcg aagaggcccg caccgatcgc
     3301 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     3361 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3421 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3481 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3541 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3601 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3661 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3721 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3781 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3841 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3901 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4141 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4321 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5281 gttcgtgcac acagcccagc ttggagcgaa cgacct
//
