LOCUS       sequence-205953-        8457 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PresentER with removable cassette for cloning of MHC-I antigen 
            (mCherry version).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8457)
  AUTHORS   Gejman RS, Chang AY, Jones HF, DiKun K, Hakimi AA, Schietinger A, 
            Scheinberg DA
  TITLE     Rejection of immunogenic tumor clones is limited by clonal fraction.
  JOURNAL   Elife. 2018 Nov 30;7. pii: 41090. doi: 10.7554/eLife.41090.
  PUBMED    30499773
REFERENCE   2  (bases 1 to 8457)
  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: "Elife."; 
            date: "2018-11-30"; volume: "7. pii: 41090. doi"; pages: " 
            10.7554/eLife.41090"
COMMENT     Sequence Label: PresentER-Cassette mCherry
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           1445..1538
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2126..2238
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        2252..2751
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2287..2310)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2684..2703
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2772..3371
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2772..2791)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3268..3288
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3432..3999
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3449..4001
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3593..3610)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3820..3839
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4013..4723
                     /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(4158..4176)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4426..4445
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4821..5335
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5528..5544
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5552..5558,5559..5576,5577..5582))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5597..5618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5735..5752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5906..6494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5986..6005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6665..7456,7457..7525))
                     /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     7288..7307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7526..7630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7698..7716
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7754..7776)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7876..7895
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8089..8111
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8239..8258
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctctc gactagggat aacagggtaa
     1441 ttgtttgaat gaggcttcag tactttacag aatcgttgcc tgcacatctt ggaaacactt
     1501 gctgggatta cttcttcagg ttaacccaac agaaggctcg aggtgactgg agttcagacg
     1561 tgtgctcttc cgatcgccac catgcctaat catcagtccg ggtcacctac cggcagttca
     1621 gacctgctcc ttgatggcaa gaaacaacga gcccatctgg cgctgaggag aaaacggcga
     1681 cgggaaatgc gcaagattaa ccgaaaggtg agaagaatga atctcgcacc cattaaagaa
     1741 aaaacagcct ggcagcacct gcaagctctg atcttcgagg cggaagaagt gttgaagact
     1801 tctcaaactc cacagacctc cctcacgctg tttctagcac tcttggccgt attggccaca
     1861 tttgcttctg acacaactgt gttcactagc aacctcaaac agacaccatg gtgcatctga
     1921 ctcctgagga gaagtctgcc gttactgccc tgtggggcaa ggtgaacgtg gatgaagttg
     1981 gtggtgaggc cctgggcagg ttggtatcaa ggttacaaga caggtttaag gagaccaata
     2041 gaaactgggc atgtggagac agagaagggc caaacaggcc aaagatcgga agagcgtcgt
     2101 gtagggatct cgccgtatca ttgaattcaa ggggctactt taggagcaat tatcttgttt
     2161 actaaaactg aataccttgc tatctctttg atacattttt acaaagctga attaaaatgg
     2221 tataaattaa atcacttttt tcaattctac cgggtagggg aggcgctttt cccaaggcag
     2281 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     2341 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     2401 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     2461 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     2521 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     2581 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     2641 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     2701 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     2761 ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     2821 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     2881 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     2941 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     3001 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     3061 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     3121 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     3181 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     3241 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     3301 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     3361 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     3421 atgcatccaa ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3481 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3541 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3601 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3661 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3721 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3781 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     3841 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     3901 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     3961 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag
     4021 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     4081 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4141 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4201 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4261 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4321 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     4381 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     4441 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     4501 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     4561 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     4621 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     4681 gggccgccac tccaccggcg gcatggacga gctgtacaag taattaatta agaattatca
     4741 agcttatcga taccgtcgac ctgcagccaa gcttatcgat aaaataaaag attttattta
     4801 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4861 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     4921 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     4981 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     5041 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     5101 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     5161 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     5221 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     5281 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta
     5341 acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc
     5401 tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta
     5461 tggacagcgc agaaagagct atagttcatt atggacagcg cagaaagagc tggggagaat
     5521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5641 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5701 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6961 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7021 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7081 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7141 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7201 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7261 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7321 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7381 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7441 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7501 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7561 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7621 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7681 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7741 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7801 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7861 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7921 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7981 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     8041 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     8101 gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     8161 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     8221 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     8281 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
     8341 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     8401 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa agggggg
//
