LOCUS Exported 5316 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Allows for transcription of TDP-43 mRNA for injection into zebrafish embryos for BiFC assays. ACCESSION . VERSION . KEYWORDS pCS2plus-TDP-43 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 bioRxiv REFERENCE 2 (bases 1 to 5316) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5316 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" promoter complement(1346..1363) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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(1702..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(2815..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 4339..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 //