LOCUS       sequence-324136-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for transcription of TDP-43 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
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1368..1502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1392..1411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1446..1465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1615..1635)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1615..1633)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1654..1670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1678..1694
                     /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)."
     promoter        complement(join(1702..1708,1709..1726,1727..1732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1747..1768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1885..1902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2056..2644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2136..2155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2815..3606,3607..3675))
                     /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     3438..3457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3676..3780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3806..4261)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3943..3964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4155..4174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4332..5316
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccgcc accatgtctg aatatattcg ggtaaccgaa
      121 gatgagaacg atgagcccat tgaaatacca tcggaagacg atgggacggt gctgctctcc
      181 acggttacag cccagtttcc aggggcgtgt gggcttcgct acaggaatcc agtgtctcag
      241 tgtatgagag gtgtccggct ggtagaagga attctgcatg ccccagatgc tggctgggga
      301 aatctggtgt atgttgtcaa ctatccaaaa gataacaaaa gaaaaatgga tgagacagat
      361 gcttcatcag cagtgaaagt gaaaagagca gtccagaaaa catccgattt aatagtgttg
      421 ggtctcccat ggaaaacaac cgaacaggac ctgaaagagt attttagtac ctttggagaa
      481 gttcttatgg tgcaggtcaa gaaagatctt aagactggtc attcaaaggg gtttggcttt
      541 gttcgtttta cggaatatga aacacaagtg aaagtaatgt cacagcgaca tatgatagat
      601 ggacgatggt gtgactgcaa acttcctaat tctaagcaaa gccaagatga gcctttgaga
      661 agcagaaaag tgtttgtggg gcgctgtaca gaggacatga ctgaggatga gctgcgggag
      721 ttcttctctc agtacgggga tgtgatggat gtcttcatcc ccaagccatt cagggccttt
      781 gcctttgtta catttgcaga tgatcagatt gcgcagtctc tttgtggaga ggacttgatc
      841 attaaaggaa tcagcgttca tatatccaat gccgaaccta agcacaatag caatagacag
      901 ttagaaagaa gtggaagatt tggtggtaat ccaggtggct ttgggaatca gggtggattt
      961 ggtaatagca gagggggtgg agctggtttg ggaaacaatc aaggtagtaa tatgggtggt
     1021 gggatgaact ttggtgcgtt cagcattaat ccagccatga tggctgccgc ccaggcagca
     1081 ctacagagca gttggggtat gatgggcatg ttagccagcc agcagaacca gtcaggccca
     1141 tcgggtaata accaaaacca aggcaacatg cagagggagc caaaccaggc cttcggttct
     1201 ggaaataact cttatagtgg ctctaattct ggtgcagcaa ttggttgggg atcagcatcc
     1261 aatgcagggt cgggcagtgg ttttaatgga ggctttggct caagcatgga ttctaagtct
     1321 tctggctggg gaatgtagtc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1381 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1501 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1561 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1621 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1681 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     1741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3781 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3841 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3901 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3961 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4021 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4081 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4141 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4201 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4261 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4321 ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata
     4381 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
     4441 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
     4501 tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
     4561 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
     4621 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
     4681 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
     4741 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
     4801 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
     4861 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
     4921 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
     4981 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
     5041 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
     5101 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
     5161 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
     5221 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
     5281 tgggaggtct atataagcaa tgctcgttta gggaac
//
