LOCUS       sequence-324299-        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarlet-Hsp70 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  AUTHORS   Adriaenssens E, Tedesco B, Mediani L, Asselbergh B, Crippa V, 
            Antoniani F, Carra S, Poletti A, Timmerman V
  TITLE     BAG3 Pro209 mutants associated with myopathy and neuropathy relocate
            chaperones of the CASA-complex to aggresomes.
  JOURNAL   Sci Rep. 2020 May 29;10(1):8755. doi: 10.1038/s41598-020-65664-z.
  PUBMED    32472079
REFERENCE   2  (bases 1 to 6606)
  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: "Sci Rep."; 
            date: "2020-05-29"; volume: "10(1):8755. doi"; pages: " 
            10.1038/s41598-020-65664-z"
COMMENT     Sequence Label: pmScarlet-HSP70
FEATURES             Location/Qualifiers
     source          1..6606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..120
                     /gene="bla"
                     /label=AmpR promoter
     promoter        122..479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        122..313
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(145..165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      330..465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             514..1308
                     /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             514..1308
                     /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(568..587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1178..1197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1496..1515)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1540..1587
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1916..2504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2405..2424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2679..2982
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2983..3186
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3137..3157
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3225..3234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3231..3926
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3364..3382)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3632..3651
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    6012..6133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6049..6068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6103..6122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6140..6595)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6277..6298)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6489..6508
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
       61 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      121 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      301 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
      481 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      541 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      601 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      661 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
      721 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      781 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      841 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      901 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      961 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1021 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     1081 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1141 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1201 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1261 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     1321 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     1501 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1561 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1621 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1681 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1741 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     1801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2581 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     2641 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2701 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2761 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2821 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2881 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2941 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     3001 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     3061 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     3121 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     3181 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
     3241 agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga
     3301 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3361 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc
     3421 ctcagttcat gtacggctcc agggccttca ccaagcaccc cgccgacatc cccgactact
     3481 ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3541 gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga
     3601 agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct
     3661 gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga
     3721 tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg
     3781 ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct
     3841 cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca
     3901 ccggcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgatgg
     3961 ccaaagccgc ggcgatcggc atcgacctgg gcaccaccta ctcctgcgtg ggggtgttcc
     4021 aacacggcaa ggtggagatc atcgccaacg accagggcaa ccgcaccacc cccagctacg
     4081 tggccttcac ggacaccgag cggctcatcg gggatgcggc caagaaccag gtggcgctga
     4141 acccgcagaa caccgtgttt gacgcgaagc ggctgattgg ccgcaagttc ggcgacccgg
     4201 tggtgcagtc ggacatgaag cactggcctt tccaggtgat caacgacgga gacaagccca
     4261 aggtgcaggt gagctacaag ggggagacca aggcattcta ccccgaggag atctcgtcca
     4321 tggtgctgac caagatgaag gagatcgccg aggcgtacct gggctacccg gtgaccaacg
     4381 cggtgatcac cgtgccggcc tacttcaacg actcgcagcg ccaggccacc aaggatgcgg
     4441 gtgtgatcgc ggggctcaac gtgctgcgga tcatcaacga gcccacggcc gccgccatcg
     4501 cctacggcct ggacagaacg ggcaaggggg agcgcaacgt gctcatcttt gacctgggcg
     4561 ggggcacctt cgacgtgtcc atcctgacga tcgacgacgg catcttcgag gtgaaggcca
     4621 cggccgggga cacccacctg ggtggggagg actttgacaa caggctggtg aaccacttcg
     4681 tggaggagtt caagagaaaa cacaagaagg acatcagcca gaacaagcga gccgtgaggc
     4741 ggctgcgcac cgcctgcgag agggccaaga ggaccctgtc gtccagcacc caggccagcc
     4801 tggagatcga ctccctgttt gagggcatcg acttctacac gtccatcacc agggcgaggt
     4861 tcgaggagct gtgctccgac ctgttccgaa gcaccctgga gcccgtggag aaggctctgc
     4921 gcgacgccaa gctggacaag gcccagattc acgacctggt cctggtcggg ggctccaccc
     4981 gcatccccaa ggtgcagaag ctgctgcagg acttcttcaa cgggcgcgac ctgaacaaga
     5041 gcatcaaccc cgacgaggct gtggcctacg gggcggcggt gcaggcggcc atcctgatgg
     5101 gggacaagtc cgagaacgtg caggacctgc tgctgctgga cgtggctccc ctgtcgctgg
     5161 ggctggagac ggccggaggc gtgatgactg ccctgatcaa gcgcaactcc accatcccca
     5221 ccaagcagac gcagatcttc accacctact ccgacaacca acccggggtg ctgatccagg
     5281 tgtacgaggg cgagagggcc atgacgaaag acaacaatct gttggggcgc ttcgagctga
     5341 gcggcatccc tccggccccc aggggcgtgc cccagatcga ggtgaccttc gacatcgatg
     5401 ccaacggcat cctgaacgtc acggccacgg acaagagcac cggcaaggcc aacaagatca
     5461 ccatcaccaa cgacaagggc cgcctgagca aggaggagat cgagcgcatg gtgcaggagg
     5521 cggagaagta caaagcggag gacgaggtgc agcgcgagag ggtgtcagcc aagaacgccc
     5581 tggagtccta cgccttcaac atgaagagcg ccgtggagga tgaggggctc aagggcaaga
     5641 tcagcgaggc ggacaagaag aaggtgctgg acaagtgtca agaggtcatc tcgtggctgg
     5701 acgccaacac cttggccgag aaggacgagt ttgagcacaa gaggaaggag ctggagcagg
     5761 tgtgtaaccc catcatcagc ggactgtacc agggtgccgg tggtcccggg cctgggggct
     5821 tcggggctca gggtcccaag ggagggtctg ggtcaggccc caccattgag gaggtagatt
     5881 agggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     5941 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     6001 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6121 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6181 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6241 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6301 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6361 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6421 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6481 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6541 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     6601 ggcact
//
