LOCUS       sequence-204034-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             601..1308
                     /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(746..764)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1014..1033
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1309..1365
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3694..3815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3731..3750)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3785..3804
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3822..4277)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3959..3980)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4171..4190
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4304..4408
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4410..4767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4410..4601
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4433..4453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4618..4753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4802..5596
                     /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             4802..5596
                     /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(4856..4875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5466..5485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5784..5803)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5828..5875
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6204..6792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6693..6712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      601 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
      661 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      781 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
      841 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      961 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1021 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1081 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1141 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1201 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1261 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagag atctcgagct
     1321 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccatgcc tgaggaaacc
     1381 cagacccaag accaaccgat ggaggaggag gaggttgaga cgttcgcctt tcaggcagaa
     1441 attgcccagt tgatgtcatt gatcatcaat actttctact cgaacaaaga gatctttctg
     1501 agagagctca tttcaaattc atcagatgca ttggacaaaa tccggtatga aagcttgaca
     1561 gatcccagta aattagactc tgggaaagag ctgcatatta accttatacc gaacaaacaa
     1621 gatcgaactc tcactattgt ggatactgga attggaatga ccaaggctga cttgatcaat
     1681 aaccttggta ctatcgccaa gtctgggacc aaagcgttca tggaagcttt gcaggctggt
     1741 gcagatatct ctatgattgg ccagttcggt gttggttttt attctgctta tttggttgct
     1801 gagaaagtaa ctgtgatcac caaacataac gatgatgagc agtacgcttg ggagtcctca
     1861 gcagggggat cattcacagt gaggacagac acaggtgaac ctatgggtcg tggaacaaaa
     1921 gttatcctac acctgaaaga agaccaaact gagtacttgg aggaacgaag aataaaggag
     1981 attgtgaaga aacattctca gtttattgga tatcccatta ctctttttgt ggagaaggaa
     2041 cgtgataaag aagtaagcga tgatgaggct gaagaaaagg aagacaaaga agaagaaaaa
     2101 gaaaaagaag agaaagagtc ggaagacaaa cctgaaattg aagatgttgg ttctgatgag
     2161 gaagaagaaa agaaggatgg tgacaagaag aagaagaaga agattaagga aaagtacatc
     2221 gatcaagaag agctcaacaa aacaaagccc atctggacca gaaatcccga cgatattact
     2281 aatgaggagt acggagaatt ctataagagc ttgaccaatg actgggaaga tcacttggca
     2341 gtgaagcatt tttcagttga aggacagttg gaattcagag cccttctatt tgtcccacga
     2401 cgtgctcctt ttgatctgtt tgaaaacaga aagaaaaaga acaatatcaa attgtatgta
     2461 cgcagagttt tcatcatgga taactgtgag gagctaatcc ctgaatatct gaacttcatt
     2521 agaggggtgg tagactcgga ggatctccct ctaaacatat cccgtgagat gttgcaacaa
     2581 agcaaaattt tgaaagttat caggaagaat ttggtcaaaa aatgcttaga actctttact
     2641 gaactggcgg aagataaaga gaactacaag aaattctatg agcagttctc taaaaacata
     2701 aagcttggaa tacacgaaga ctctcaaaat cggaagaagc tttcagagct gttaaggtac
     2761 tacacatctg cctctggtga tgagatggtt tctctcaagg actactgcac cagaatgaag
     2821 gagaaccaga aacatatcta ttatatcaca ggtgagacca aggaccaggt agctaactca
     2881 gcctttgtgg aacgtcttcg gaaacatggc ttagaagtga tctatatgat tgagcccatt
     2941 gatgagtact gtgtccaaca gctgaaggaa tttgagggga agactttagt gtcagtcacc
     3001 aaagaaggcc tggaacttcc agaggatgaa gaagagaaaa agaagcagga agagaaaaaa
     3061 acaaagtttg agaacctctg caaaatcatg aaagacatat tggagaaaaa agttgaaaag
     3121 gtggttgtgt caaaccgatt ggtgacatct ccatgctgta ttgtcacaag cacatatggc
     3181 tggacagcaa acatggagag aatcatgaaa gctcaagccc taagagacaa ctcaacaatg
     3241 ggttacatgg cagcaaagaa acacctggag ataaaccctg accattccat tattgagacc
     3301 ttaaggcaaa aggcagaggc tgataagaac gacaagtctg tgaaggatct ggtcatcttg
     3361 ctttatgaaa ctgcgctcct gtcttctggc ttcagtctgg aagatcccca gacacatgct
     3421 aacaggatct acaggatgat caaacttggt ctgggtattg atgaagatga ccctactgct
     3481 gatgatacca gtgctgctgt aactgaagaa atgccacccc ttgaaggaga tgacgacaca
     3541 tcacgcatgg aagaagtaga ctaaggatcc accggatcta gataactgat cataatcagc
     3601 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3661 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3721 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3781 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3841 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3901 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3961 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4021 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4081 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4141 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4201 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4261 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4321 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4381 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4441 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4561 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4621 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4681 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4741 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4801 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4861 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4921 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4981 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5041 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5101 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5161 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5221 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5281 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5341 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5401 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5461 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5521 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5581 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5641 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5701 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5761 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5821 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5881 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5941 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6001 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6061 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6901 atgcat
//
