LOCUS       sequence-205952-        8312 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PresentER minigene encoding H2-Kb ligand SIINFEKL (mCherry).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8312)
  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 8312)
  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-SIINFEKL (mCherry)
FEATURES             Location/Qualifiers
     source          1..8312
                     /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           1979..2091
                     /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        2105..2604
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2140..2163)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2537..2556
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2625..3224
                     /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(2625..2644)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3121..3141
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3285..3852
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3302..3854
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3446..3463)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3673..3692
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3866..4576
                     /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(4011..4029)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4279..4298
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4674..5188
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5383..5399
                     /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(5407..5413,5414..5431,5432..5437))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5452..5473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5590..5607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5761..6349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5841..5860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6520..7311,7312..7380))
                     /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     7143..7162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7381..7485)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7553..7571
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7609..7631)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7731..7750
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7944..7966
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8094..8113
                     /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 attggccccg
     1861 ccacctgtga gcgggagcat catcaacttc gagaagctgt aaggccaaac aggccaaaga
     1921 tcggaagagc gtcgtgtagg gaaagagtgt agatctcggt ggtcgccgta tcattgaatt
     1981 caaggggcta ctttaggagc aattatcttg tttactaaaa ctgaatacct tgctatctct
     2041 ttgatacatt tttacaaagc tgaattaaaa tggtataaat taaatcactt ttttcaattc
     2101 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     2161 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     2221 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     2281 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     2341 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     2401 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     2461 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     2521 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     2581 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     2641 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     2701 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     2761 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     2821 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     2881 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     2941 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     3001 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     3061 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     3121 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     3181 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     3241 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc caattccgcc ccccccccct
     3301 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3361 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3421 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3481 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3541 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3601 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3661 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3721 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3781 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3841 acacgatgat aatatggcca caaccatggt gagcaagggc gaggaggata acatggccat
     3901 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     3961 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     4021 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     4081 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     4141 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     4201 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     4261 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     4321 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     4381 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     4441 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     4501 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     4561 cgagctgtac aagtaattaa ttaagaatta tcaagcttat cgataccgtc gacctgcagc
     4621 caagcttatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     4681 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     4741 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     4801 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4861 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     4921 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4981 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     5041 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct
     5101 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     5161 tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca
     5221 ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc
     5281 cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga
     5341 attgtgaagt tagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     5401 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5461 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5521 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5581 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5641 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7261 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7381 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7441 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7501 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7561 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     7621 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     7681 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     7741 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     7801 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     7861 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     7921 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     7981 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     8041 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     8101 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     8161 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     8221 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     8281 aagattttat ttagtctcca gaaaaagggg gg
//
