LOCUS       sequence-205954-        8468 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PresentER with removable cassette for cloning of MHC-I antigen (GFP 
            version).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8468)
  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 8468)
  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 GFP
FEATURES             Location/Qualifiers
     source          1..8468
                     /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           2122..2234
                     /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        2248..2747
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2283..2306)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2680..2699
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2768..3367
                     /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(2768..2787)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3264..3284
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3428..3995
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3445..3997
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3589..3606)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3816..3835
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4009..4011,4012..4014,4015..4728)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4054..4075)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4315..4334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4662..4683
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4830..5344
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5539..5555
                     /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(5563..5569,5570..5587,5588..5593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5608..5629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5746..5763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5917..6505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5997..6016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6676..7467,7468..7536))
                     /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     7299..7318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7537..7641)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7709..7727
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7765..7787)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7887..7906
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8100..8122
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8250..8269
                     /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 gtagggaacc gtatcattga attcaagggg ctactttagg agcaattatc ttgtttacta
     2161 aaactgaata ccttgctatc tctttgatac atttttacaa agctgaatta aaatggtata
     2221 aattaaatca cttttttcaa ttctaccggg taggggaggc gcttttccca aggcagtctg
     2281 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     2341 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     2401 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     2461 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     2521 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     2581 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     2641 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     2701 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa
     2761 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     2821 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     2881 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     2941 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3001 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3061 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     3121 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     3181 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3241 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     3301 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     3361 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     3421 atccaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     3481 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     3541 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     3601 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     3661 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     3721 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     3781 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     3841 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     3901 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3961 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     4021 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4081 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     4141 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4201 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4261 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     4321 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4381 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4441 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     4501 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     4561 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     4621 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4681 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     4741 gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata aaataaaaga
     4801 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     4861 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     4921 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     4981 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     5041 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     5101 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     5161 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     5221 tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     5281 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     5341 tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt
     5401 aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat
     5461 agttcattat ggacagcgca gaaagagctg gagagttcat tatggacagc gcagaaagag
     5521 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5581 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5641 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5701 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7681 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     7741 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7801 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     7861 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7921 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     7981 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8041 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8101 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     8161 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     8221 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     8281 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     8341 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     8401 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     8461 aagggggg
//
