LOCUS       sequence-317647-        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"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             650..1345
                     /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(783..801)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1051..1070
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3431..3552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3468..3487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3522..3541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3559..4014)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3696..3717)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3908..3927
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4041..4145
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4147..4504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4147..4338
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4170..4190)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4355..4490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4539..5333
                     /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             4539..5333
                     /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(4593..4612)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5203..5222
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5521..5540)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5565..5612
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5941..6529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6430..6449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
      661 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
      721 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      781 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
      841 tcagttcatg tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta
      901 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      961 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     1021 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     1081 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     1141 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     1201 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     1261 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     1321 cggcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgatggc
     1381 caaagccgcg gcgatcggca tcgacctggg caccacctac tcctgcgtgg gggtgttcca
     1441 acacggcaag gtggagatca tcgccaacga ccagggcaac cgcaccaccc ccagctacgt
     1501 ggccttcacg gacaccgagc ggctcatcgg ggatgcggcc aagaaccagg tggcgctgaa
     1561 cccgcagaac accgtgtttg acgcgaagcg gctgattggc cgcaagttcg gcgacccggt
     1621 ggtgcagtcg gacatgaagc actggccttt ccaggtgatc aacgacggag acaagcccaa
     1681 ggtgcaggtg agctacaagg gggagaccaa ggcattctac cccgaggaga tctcgtccat
     1741 ggtgctgacc aagatgaagg agatcgccga ggcgtacctg ggctacccgg tgaccaacgc
     1801 ggtgatcacc gtgccggcct acttcaacga ctcgcagcgc caggccacca aggatgcggg
     1861 tgtgatcgcg gggctcaacg tgctgcggat catcaacgag cccacggccg ccgccatcgc
     1921 ctacggcctg gacagaacgg gcaaggggga gcgcaacgtg ctcatctttg acctgggcgg
     1981 gggcaccttc gacgtgtcca tcctgacgat cgacgacggc atcttcgagg tgaaggccac
     2041 ggccggggac acccacctgg gtggggagga ctttgacaac aggctggtga accacttcgt
     2101 ggaggagttc aagagaaaac acaagaagga catcagccag aacaagcgag ccgtgaggcg
     2161 gctgcgcacc gcctgcgaga gggccaagag gaccctgtcg tccagcaccc aggccagcct
     2221 ggagatcgac tccctgtttg agggcatcga cttctacacg tccatcacca gggcgaggtt
     2281 cgaggagctg tgctccgacc tgttccgaag caccctggag cccgtggaga aggctctgcg
     2341 cgacgccaag ctggacaagg cccagattca cgacctggtc ctggtcgggg gctccacccg
     2401 catccccaag gtgcagaagc tgctgcagga cttcttcaac gggcgcgacc tgaacaagag
     2461 catcaacccc gacgaggctg tggcctacgg ggcggcggtg caggcggcca tcctgatggg
     2521 ggacaagtcc gagaacgtgc aggacctgct gctgctggac gtggctcccc tgtcgctggg
     2581 gctggagacg gccggaggcg tgatgactgc cctgatcaag cgcaactcca ccatccccac
     2641 caagcagacg cagatcttca ccacctactc cgacaaccaa cccggggtgc tgatccaggt
     2701 gtacgagggc gagagggcca tgacgaaaga caacaatctg ttggggcgct tcgagctgag
     2761 cggcatccct ccggccccca ggggcgtgcc ccagatcgag gtgaccttcg acatcgatgc
     2821 caacggcatc ctgaacgtca cggccacgga caagagcacc ggcaaggcca acaagatcac
     2881 catcaccaac gacaagggcc gcctgagcaa ggaggagatc gagcgcatgg tgcaggaggc
     2941 ggagaagtac aaagcggagg acgaggtgca gcgcgagagg gtgtcagcca agaacgccct
     3001 ggagtcctac gccttcaaca tgaagagcgc cgtggaggat gaggggctca agggcaagat
     3061 cagcgaggcg gacaagaaga aggtgctgga caagtgtcaa gaggtcatct cgtggctgga
     3121 cgccaacacc ttggccgaga aggacgagtt tgagcacaag aggaaggagc tggagcaggt
     3181 gtgtaacccc atcatcagcg gactgtacca gggtgccggt ggtcccgggc ctgggggctt
     3241 cggggctcag ggtcccaagg gagggtctgg gtcaggcccc accattgagg aggtagatta
     3301 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3361 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3421 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3541 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3601 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3661 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3721 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3781 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3841 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3901 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3961 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4021 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4081 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4141 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4201 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4261 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4321 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4381 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4441 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4501 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4561 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4621 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4681 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4741 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4801 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4861 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4921 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4981 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5041 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5101 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5161 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5221 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5281 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5341 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5401 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5461 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5521 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5581 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5641 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5701 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5761 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5821 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5881 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5941 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6001 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6061 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6121 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6181 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6241 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6301 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6361 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6421 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6481 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6541 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6601 gttatc
//
