LOCUS       sequence-207849-        6906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for mCherry fusion to human Hsp90 alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  AUTHORS   Lev A, Takeda K, Zanker D, Maynard JC, Dimberu P, Waffarn E, Gibbs 
            J, Netzer N, Princiotta MF, Neckers L, Picard D, Nicchitta CV, Chen 
            W, Reiter Y, Bennink JR, Yewdell JW
  TITLE     The exception that reinforces the rule: crosspriming by cytosolic 
            peptides that escape degradation.
  JOURNAL   Immunity. 2008 Jun;28(6):787-98. doi: 10.1016/j.immuni.2008.04.015.
  PUBMED    18549799
REFERENCE   2  (bases 1 to 6906)
  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"; doi: 
            "10.1016/j.immuni.2008.04"; journalName: "Immunity"; date: 
            "2008-06"; volume: "28"; issue: "6"; pages: "787-98"
COMMENT     Sequence Label: pCherry.90 alpha
FEATURES             Location/Qualifiers
     source          1..6906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        116..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        420..623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     574..594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             656..1363
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(801..819)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1069..1088
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1364..1420
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3749..3870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3786..3805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3840..3859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3877..4332)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4014..4035)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4226..4245
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4359..4463
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4465..4822
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4465..4656
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4488..4508)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4673..4808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4857..5651
                     /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             4857..5651
                     /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(4911..4930)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5521..5540
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5839..5858)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5883..5930
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6259..6847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6748..6767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      601 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag ccaccatggt
      661 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
      721 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      781 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
      841 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
      901 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      961 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1021 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1081 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1141 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1201 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1261 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1321 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagagatctc gagctcaagc
     1381 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc atgcctgagg aaacccagac
     1441 ccaagaccaa ccgatggagg aggaggaggt tgagacgttc gcctttcagg cagaaattgc
     1501 ccagttgatg tcattgatca tcaatacttt ctactcgaac aaagagatct ttctgagaga
     1561 gctcatttca aattcatcag atgcattgga caaaatccgg tatgaaagct tgacagatcc
     1621 cagtaaatta gactctggga aagagctgca tattaacctt ataccgaaca aacaagatcg
     1681 aactctcact attgtggata ctggaattgg aatgaccaag gctgacttga tcaataacct
     1741 tggtactatc gccaagtctg ggaccaaagc gttcatggaa gctttgcagg ctggtgcaga
     1801 tatctctatg attggccagt tcggtgttgg tttttattct gcttatttgg ttgctgagaa
     1861 agtaactgtg atcaccaaac ataacgatga tgagcagtac gcttgggagt cctcagcagg
     1921 gggatcattc acagtgagga cagacacagg tgaacctatg ggtcgtggaa caaaagttat
     1981 cctacacctg aaagaagacc aaactgagta cttggaggaa cgaagaataa aggagattgt
     2041 gaagaaacat tctcagttta ttggatatcc cattactctt tttgtggaga aggaacgtga
     2101 taaagaagta agcgatgatg aggctgaaga aaaggaagac aaagaagaag aaaaagaaaa
     2161 agaagagaaa gagtcggaag acaaacctga aattgaagat gttggttctg atgaggaaga
     2221 agaaaagaag gatggtgaca agaagaagaa gaagaagatt aaggaaaagt acatcgatca
     2281 agaagagctc aacaaaacaa agcccatctg gaccagaaat cccgacgata ttactaatga
     2341 ggagtacgga gaattctata agagcttgac caatgactgg gaagatcact tggcagtgaa
     2401 gcatttttca gttgaaggac agttggaatt cagagccctt ctatttgtcc cacgacgtgc
     2461 tccttttgat ctgtttgaaa acagaaagaa aaagaacaat atcaaattgt atgtacgcag
     2521 agttttcatc atggataact gtgaggagct aatccctgaa tatctgaact tcattagagg
     2581 ggtggtagac tcggaggatc tccctctaaa catatcccgt gagatgttgc aacaaagcaa
     2641 aattttgaaa gttatcagga agaatttggt caaaaaatgc ttagaactct ttactgaact
     2701 ggcggaagat aaagagaact acaagaaatt ctatgagcag ttctctaaaa acataaagct
     2761 tggaatacac gaagactctc aaaatcggaa gaagctttca gagctgttaa ggtactacac
     2821 atctgcctct ggtgatgaga tggtttctct caaggactac tgcaccagaa tgaaggagaa
     2881 ccagaaacat atctattata tcacaggtga gaccaaggac caggtagcta actcagcctt
     2941 tgtggaacgt cttcggaaac atggcttaga agtgatctat atgattgagc ccattgatga
     3001 gtactgtgtc caacagctga aggaatttga ggggaagact ttagtgtcag tcaccaaaga
     3061 aggcctggaa cttccagagg atgaagaaga gaaaaagaag caggaagaga aaaaaacaaa
     3121 gtttgagaac ctctgcaaaa tcatgaaaga catattggag aaaaaagttg aaaaggtggt
     3181 tgtgtcaaac cgattggtga catctccatg ctgtattgtc acaagcacat atggctggac
     3241 agcaaacatg gagagaatca tgaaagctca agccctaaga gacaactcaa caatgggtta
     3301 catggcagca aagaaacacc tggagataaa ccctgaccat tccattattg agaccttaag
     3361 gcaaaaggca gaggctgata agaacgacaa gtctgtgaag gatctggtca tcttgcttta
     3421 tgaaactgcg ctcctgtctt ctggcttcag tctggaagat ccccagacac atgctaacag
     3481 gatctacagg atgatcaaac ttggtctggg tattgatgaa gatgacccta ctgctgatga
     3541 taccagtgct gctgtaactg aagaaatgcc accccttgaa ggagatgacg acacatcacg
     3601 catggaagaa gtagactaag gatccaccgg atctagataa ctgatcataa tcagccatac
     3661 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3721 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3781 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3841 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     3901 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3961 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4021 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4081 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4141 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4201 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4261 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4321 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4381 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4441 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4501 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4561 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4621 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4681 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4741 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4801 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4861 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4921 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4981 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5041 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5101 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5161 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5221 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5281 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5341 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5401 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5461 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5521 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5581 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5641 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5701 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5761 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5821 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5881 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5941 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6001 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6061 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6121 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6241 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6301 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6361 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6421 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6481 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6541 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6601 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6661 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6721 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6781 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6841 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6901 tcacat
//
