LOCUS       sequence-495984-        5865 bp    DNA     circular SYN 23-DEC-2025
DEFINITION  This plasmid expresses the mRuby3 flourescent tag for integration 
            into mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  AUTHORS   Gracia B, Zhang XH, Montes P, Pham TC, Huang M, Chen J, Karras GI
  TITLE     HSP90 buffers deleterious genetic variations in BRCA1.
  JOURNAL   Mol Cell. 2025 Nov 19:S1097-2765(25)00867-6. doi: 
            10.1016/j.molcel.2025.10.026.
  PUBMED    41265458
REFERENCE   2  (bases 1 to 5865)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2025 Nov 19:S1097-2765(25)00867-6. doi: 
            10.1016/j.molcel.2025.10.026."
COMMENT     Sequence Label: pKlab639-HBEGF-2a-mRuby3-WPRE
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..78)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(171..759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(251..270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(930..1721,1722..1790))
                     /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     1553..1572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1791..1895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2196..2218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2210..2227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2211..2227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2261..2280
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2261..2279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3371..3424
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3437..4144
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     misc_feature    4165..4753
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4218..4238)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4636..4647)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4791..4808)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4797..5021
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc
       61 ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca
      121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc
     1081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1921 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1981 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2041 tagggttgag tggccgctac agggcgctcc cattcgccat tcaggctgcg caactgttgg
     2101 gaagggcgtt tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     2161 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     2221 ggccagtccc ccggtcaggc aacatcaccc gagcgcgacg taatacgact cactataggg
     2281 cgaattggcg gaaggccgtc aaggccgcat acgcgtccat gtcttcggaa atacaaggac
     2341 ttctgcatcc atggagaatg caaatatgtg aaggagctcc gggctccctc ctgcatgtaa
     2401 gtgccccttc cccagggctg aatctcatca gcacactttg tcagccacgt ggctgttcct
     2461 cgttgtcact gttccttgaa ttcatgattt cacccagttt cttctcaacc tctgggcgga
     2521 agttgggagg aggggaaata tatttttagt cagcggaagc cccctccccc ctataggatg
     2581 caatttcctg tggtatggtt ttgtgacgtg ctttaatcct tggggacatt tcctgcttgc
     2641 ccagaaatga gcatgtggct aggacagctg gcacctgaag gcaggccctt aattcttgcc
     2701 tgatgcccta ctctgggagg gagaagccag taggaaacat ggcagagtgg gcttccaggg
     2761 cagagtagag ctcctgtggg aaggtaggaa gtgcatttgg atgcatgatg tataggtatg
     2821 tgtgtatttg ggtttatgtg catgtaagtg tgcaaatgtg gattgactgt gaggcatggc
     2881 aggactgtac agagagggat catcatggcg gcaggttgag gcctctcttt cttcttcctt
     2941 atcccagcaa ggacgaggag gtgggagaca tggagagtac tggcctttgg ccacgttgtg
     3001 agagaacaat tcctttgtgc agggttcaca ggaaatggaa cctgacccat taggcatcag
     3061 cccccggaat tctagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
     3121 tccctttttt ttccacagct gccacccggg ttaccatgga aaaaggtgtc atgggctgag
     3181 cctcccagtg gaaaatcgct tatataccta tgaccacaca accatcctgg ccgtggtggc
     3241 tgtggtgctg tcatctgtct gtctgctggt catcgtgggg cttctcatgt ttaggtacca
     3301 taggagagga ggttatgatg tggaaaatga agagaaagtg aagttgggca tgactaattc
     3361 ccacggatcc gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     3421 cccagcaccg ggatccgtgt ctaagggcga agagctgatc aaggaaaata tgcgtatgaa
     3481 ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg
     3541 cagaccgtac gagggagtgc aaaccatgag gatcaaagtc atcgagggag gacccctgcc
     3601 atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtacct ttatcaagta
     3661 cccggccgac atccctgatt tctttaaaca gtcctttcct gagggtttta cttgggaaag
     3721 agttacgaga tacgaagatg gtggagtcgt caccgtcacg caggacacca gccttgagga
     3781 tggcgagctc gtctacaacg tcaaggtcag aggggtaaac tttccctcca atggtcccgt
     3841 gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc cagcagatgg
     3901 tggtctgaga ggatacactg acatcgcact gaaagttgat ggtggtggcc atctgcactg
     3961 caacttcgtg acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtgt
     4021 ccatgccgtt gatcaccgcc tggaaaggat cgaggagagt gacaatgaaa cctacgtagt
     4081 gcaaagagaa gtggcagttg ccaaatacag caaccttggt ggtggcatgg acgagctgta
     4141 caagtaatag tgatgacctg caggaatcaa cctctggatt acaaaatttg tgaaagattg
     4201 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4261 ttgtatcatg ctattgcttc ccgtatggcc ttcattttct cctccttgta taaatcctgg
     4321 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4381 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4441 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4501 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     4561 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4621 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4681 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4741 gccgcctccc cgcctgtaat ctagatagtc agtttaaacc cgctgatcag cctcgactgt
     4801 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4861 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4921 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg gggattggga
     4981 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctctcg aggtcaggca
     5041 acatcacccc ttccctgggt aggtgtgtgg gtggaggggc tgtgggttcc ttagcctctc
     5101 tcctaagcca aacccagcaa acggctgcct tggcaacccc tcagggatga cagcactgcc
     5161 atgctctctg gcaggcataa tgttgccact gtgcctgagg ccaacaccct gcgtcgggct
     5221 gcaaacatcc attcccttcc ctgtggggag ggaggctctg ggggccttag tgggagactc
     5281 tggacagggc caagagactg ttgtatgcac actgcctcca gcctgtcaag aaggcggcgt
     5341 gcctggcatc ccttctactg gtgattggtg cagatccctt agctttttaa agcttccttg
     5401 ttttgtctga tcacacacag cagagctgcc ctgtatttgg cagttggcag acagacccat
     5461 cactccccac catgtccaca gtcacttgtg catcctttcc tataacatcc ttgtcaggag
     5521 cttggtatta gagggagttg tttaagagtg gcatagaaag cccccatatt atccttccca
     5581 aggtcttggg acagggtggg aaatgttcat cttaaatttg taaaatggca tcattagtac
     5641 agggtgaaga aggtgactca agtagtcaag gtggattgag gtcaggaatc tgtctatacc
     5701 agattggtcc tgggcatttt ggtggatgga tgtggggctt gcactgtgtg gttgagaggc
     5761 cttataaggt tgccctcctg gaggtaccct gggcctcatg ggccttccgc tcactgcccg
     5821 ctttccagtc gggaaacctg tccccccggt caggcaacat caccc
//
