LOCUS       sequence-364093-        6213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tandem MS2 binding protein labeled with tdTomato, containing a NLS 
            and a HA-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 6213)
  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"; journalName: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: NLS-HA-tdMCP-tdTomato
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             643..663
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             667..693
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             667..693
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             694..705
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1450..1459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1456..2886
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3045..3064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3099..3118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3136..3591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3273..3294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3485..3504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3618..3722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3724..4081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3724..3915
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3747..3767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3932..4067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4116..4910
                     /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             4116..4910
                     /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(4170..4189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4780..4799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5098..5117)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5142..5189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5518..6106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6007..6026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgaca
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc ggggggggca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gctttcatgg gcccaaaaaa gaaaagaaaa
      661 gttggctacc cctacgacgt gcccgactac gccatcgaag gccgccatat gctagccgtt
      721 aaaatggctt ctaactttac tcagttcgtt ctcgtcgaca atggcggaac tggcgacgtg
      781 actgtcgccc caagcaactt cgctaacggg atcgctgaat ggatcagctc taactcgcgt
      841 tcacaggctt acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa tcgcaaatac
      901 accatcaaag tcgaggtgcc taaaggcgcc tggcgttcgt acttaaatat ggaactaacc
      961 attccaattt tcgccacgaa ttccgactgc gagcttattg ttaaggcaat gcaaggtctc
     1021 ctaaaagatg gaaacccgat tccctcagca atcgcagcaa actccggcat ctacgccatg
     1081 gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga cgtgactgtc
     1141 gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc gcgttcacag
     1201 gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa atacaccatc
     1261 aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact aaccattcca
     1321 attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg tctcctaaaa
     1381 gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgc ggattctaga
     1441 gaaccggtcg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc
     1501 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     1561 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     1621 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
     1681 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
     1741 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
     1801 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
     1861 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     1921 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
     1981 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
     2041 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2101 gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc
     2161 ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg
     2221 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     2281 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     2341 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     2401 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     2461 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     2521 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     2581 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     2641 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     2701 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     2761 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     2821 cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac
     2881 aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2941 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3001 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3121 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3181 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3241 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3301 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3361 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3421 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3481 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3541 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3721 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3961 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4021 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4081 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4141 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4201 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4261 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4321 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4381 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4441 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4501 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4561 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4621 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4681 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4741 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4801 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4861 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4921 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4981 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5041 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5101 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5161 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5221 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5281 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5341 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5641 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5881 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgc
//
