LOCUS       sequence-270554-        7256 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged HLA-A*02:01 (I52P 
            mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7256)
  AUTHORS   McShan AC, Devlin CA, Overall SA, Park J, Toor JS, Moschidi D, 
            Flores-Solis D, Choi H, Tripathi S, Procko E, Sgourakis NG
  TITLE     Molecular determinants of chaperone interactions on MHC-I for 
            folding and antigen repertoire selection.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Dec 17;116(51):25602-25613. doi: 
            10.1073/pnas.1915562116. Epub 2019 Dec 3.
  PUBMED    31796585
REFERENCE   2  (bases 1 to 7256)
  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: 
            "10.1073/pnas.1915562116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-12-17- 17"; volume: "116"; issue: "51"; pages: 
            "25602-25613"
COMMENT     Sequence Label: pCEP4-myc-HLA-A2-VN (I52P)
FEATURES             Location/Qualifiers
     source          1..7256
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(312..330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        398..502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(503..571,572..1363)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(721..740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1534..2122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2023..2042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2553..2698
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2741..3778
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(3776..3795)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3820..3868
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4522..4901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4902..5105
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5056..5076
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5102..5126
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5208..5237
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6285..6287,6288..6746)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(6327..6348)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6588..6607)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    6769..6903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6793..6812)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6847..6866
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7046..7068)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acagatccag gtctgaaaat tccccatcct ccgaaccatc ctcgtcctca tcaccaatta
       61 ctcgcagccc ggaaaactcc cgctgaacat cctcaagatt tgcgtcctga gcctcaagcc
      121 aggcctcaaa ttcctcgtcc ccctttttgc tggacggtag ggatggggat tctcgggacc
      181 cctcctcttc ctcttcaagg tcaccagaca gagatgctac tggggcaacg gaagaaaagc
      241 tgggtgcggc ctgtgaggat cagcttatcg atgataagct gtcaaacatg agaattcttg
      301 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      361 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1621 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1681 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1741 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1801 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1861 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1921 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1981 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2041 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2101 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2161 ttgctggcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     2221 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     2281 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccccga gatgcgccgc
     2341 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     2401 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     2461 aggtgccgcc ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg
     2521 aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc agcgtcttgt
     2581 cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtccgagg tccacttcgc
     2641 atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaca
     2701 gcgtcaacag cgtgccgcag atcccggggg gcaatgagat atgaaaaagc ctgaactcac
     2761 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     2821 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     2881 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     2941 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     3001 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3061 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     3121 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3181 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3241 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     3301 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     3361 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     3421 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     3481 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     3541 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     3601 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     3661 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     3721 tagtggaaac cgacgcccca gcactcgtcc ggatcgggag atgggggagg ctaactgaaa
     3781 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     3841 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     3901 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     3961 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4021 ggccctgcca tagccactgg ccccgtgggt tagggacggg gtcccccatg gggaatggtt
     4081 tatggttcgt gggggttatt attttgggcg ttgcgtgggg tcaggtccac gactggactg
     4141 agcagacaga cccatggttt ttggatggcc tgggcatgga ccgcatgtac tggcgcgaca
     4201 cgaacaccgg gcgtctgtgg ctgccaaaca cccccgaccc ccaaaaacca ccgcgcggat
     4261 ttctggcgtg ccaagctagt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     4321 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     4381 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     4441 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     4501 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
     4561 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4621 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4681 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4741 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
     4801 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
     4861 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     4921 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4981 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
     5041 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     5101 gagctcgttt agtgaaccgt cagatctcta gaagctgggt accagctgct agcgccacca
     5161 tgaagaccat catcgccctg agctacatct tctgcctggt gttcgccgag cagaaattga
     5221 tcagcgagga ggacctgggc agccctggag gcagttcagg aggcggatct cactccatga
     5281 ggtatttctt cacatccgtg tcccggcccg gccgcgggga gccccgcttc atcgcagtgg
     5341 gctacgtgga cgacacgcag ttcgtgcggt tcgacagcga cgccgcgagc cagaggatgg
     5401 agccgcgggc gccgtggcca gagcaggagg gtccggagta ttgggacggg gagacacgga
     5461 aagtgaaggc ccactcacag actcaccgag tggacctggg gaccctgcgc ggctactaca
     5521 accagagcga ggccggttct cacaccgtcc agaggatgta tggctgcgac gtggggtcgg
     5581 actggcgctt cctccgcggg taccaccagt acgcctacga cggcaaggat tacatcgccc
     5641 tgaaagagga cctgcgctct tggaccgcgg cggacatggc agctcagacc accaagcaca
     5701 agtgggaggc ggcccatgtg gcggagcagt tgagagccta cctggagggc acgtgcgtgg
     5761 agtggctccg cagatacctg gagaacggga aggagacgct gcagcgtacg gacgccccca
     5821 aaacgcatat gactcaccac gctgtctctg accatgaagc caccctgagg tgctgggccc
     5881 tgagcttcta ccctgcggag atcacactga cctggcagcg ggatggggag gaccagaccc
     5941 aggacacgga gctcgtggag accaggcctg caggggatgg aaccttccag aagtgggcgg
     6001 ctgtggtggt gccttctgga caggagcaga gatacacctg ccatgtgcag catgagggtt
     6061 tgcccaagcc cctcaccctg agatgggagc cgtcttccca gcccaccatc cccatcgtgg
     6121 gcatcattgc tggcctggtt ctctttggag ctgtgatcac tggagctgtg gtcgctgctg
     6181 tgatgtggag gaggaagagc tcagatagaa aaggagggag ctactctcag gctgcaagca
     6241 gtgacagtgc ccagggctct gatgtgtctc tcacagcttg taaagtgagc aagggcgagg
     6301 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     6361 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     6421 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     6481 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     6541 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     6601 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     6661 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     6721 acagccacaa cgtctatctg accgcctaac tcgaggccgg caaggccgga tccagacatg
     6781 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     6841 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6901 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     6961 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg
     7021 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7081 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7141 gcgggtgtcg gggcgcagcc atgaggtcga ctctagagga tcgatccccg ccccggacga
     7201 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcc
//
