LOCUS       sequence-203558-        5680 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To test the effect of sequence on TDP43 droplet properties.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  AUTHORS   Schmidt HB, Barreau A, Rohatgi R
  TITLE     Phase separation-deficient TDP43 remains functional in splicing.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
  PUBMED    31653829
REFERENCE   2  (bases 1 to 5680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: " 
            10.1038/s41467-019-12740-2"
COMMENT     Sequence Label: pHBS1039 TDP43 RRM-GFP 2x-Phi
FEATURES             Location/Qualifiers
     source          1..5680
                     /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"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      151..160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1660..1684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1729..1863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1753..1772)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1807..1826
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1976..1996)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1976..1994)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2015..2031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2015..2031)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2028..2050)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2039..2055
                     /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(2063..2069,2070..2087,2088..2093))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2246..2263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2417..3005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2497..2516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3176..3967,3968..4036))
                     /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     3799..3818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4037..4141)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4167..4622)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4325)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4516..4535
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4696..5680
                     /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 atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctggcgccgg taccaccggt gccaccatgg gatccatgtc tgaatatatt
      181 cgggtaaccg aagatgagaa cgatgagccc attgaaatac catcggaaga cgatgggacg
      241 gtgctgctct ccacggttac agcccagttt ccaggggcgt gtgggcttcg ctacaggaat
      301 ccagtgtctc agtgtatgag aggtgtccgg ctggtagaag gaattctgca tgccccagat
      361 gctggctggg gaaatctggt gtatgttgtc aactatccaa aagataacaa aagaaaaatg
      421 gatgagacag atgcttcatc agcagtgaaa gtgaaaagag cagtccagaa aacatccagt
      481 aaaggagagg aactatttac aggtactgtg cctatcaaag tcgagctgga cggcgacgta
      541 aacggccaca agttcagcgt gcgtggcgag ggcgagggcg atgccaccga gggcaagctg
      601 accctgaagt tcatctgcac caccggcaag ctgccggtgc cgtggccgac cctcgtgacc
      661 accctgacct acggcgtgca gtgcttcagc cgctacccgg accacatgaa gcgtcacgac
      721 ttcttcaagt ccgccatgcc ggaaggctac gtccaggagc gcaccatcga gttcaaggac
      781 gacggcacct acaagacccg cgcggaggtg aagttcgagg gcgacaccct ggtgaaccgc
      841 atcgagctga agggcaacga cttcaaggag gacggcaaca tcctgggtca caagctggag
      901 tacaaccaca acagccacaa cgtccgtatc gaagccgaca agcagaagaa cggcatcaag
      961 gctaacttca agatccgcca caacgttgag gacggcagcc agcaggaagc cgaccacaaa
     1021 cagcagaaca ccccgatcgg cgacggcccg gtgcgcctgc cggacaacca ctatctgagc
     1081 acccagacca cgctgagcaa agacccgaac gagaagcgcg atcacatggt cctgaaggag
     1141 ttcgtgacgg cagccggtat cactaaaggt gaagatgaaa gtggatccaa tgccgaacct
     1201 aaacataata gcaaccgaca actcgagaga tctggtagag gtaatccgaa tcagaattcc
     1261 cgaggaggat ttgggggagg gtttggaggc gggttcggcc ttgggatggg cttcggtttt
     1321 tctggagcgg gcaacaatca gggctcaaac ggtggtatga acgcaataaa cccggctatg
     1381 atggctgcgg ctcaggccgc actacagagc agttggggaa tgatgggcat gctggctagt
     1441 cagcagaatc agagtggtcc gtcagggaac aaccagaacc agggaaatat gcagcgcgag
     1501 cctaatcagg ctagcggaaa caattcattt ggttattcaa tcggatgggg gggctttggg
     1561 ggatttggag ggtggggcat gtcaggcagt aatagcggag ccgcgagcgc ttccaatgca
     1621 gggtcaggtt cgaattctat ggacagcaaa agttcgtaga cccagctttc ttgtacaaag
     1681 tggttgatcc tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
     1741 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1801 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1861 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1921 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     1981 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2041 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2101 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2161 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2221 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2281 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2341 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2821 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3121 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3181 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3241 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3301 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3361 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3421 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3481 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3541 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3601 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3661 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3721 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3781 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3841 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3901 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3961 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4021 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4081 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4141 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4201 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4261 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4321 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4381 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4441 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4501 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4561 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4621 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4681 gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc atgccaattc
     4741 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
     4801 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
     4861 aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
     4921 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
     4981 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
     5041 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
     5101 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
     5161 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
     5221 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
     5281 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
     5341 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
     5401 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
     5461 agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc agtactccca
     5521 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
     5581 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     5641 ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
