LOCUS       sequence-270552-        9007 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged HLA-A*02:01 (V67P 
            mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9007)
  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 9007)
  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 (V67P)
FEATURES             Location/Qualifiers
     source          1..9007
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1187..1205)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1273..1377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1378..1446,1447..2238)
                     /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(1596..1615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2409..2997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2898..2917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3428..3573
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3616..4653
                     /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(4651..4670)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4695..4743
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5397..5776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5777..5980
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5931..5951
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5977..6001
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6083..6112
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(7160..7162,7163..7621)
                     /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(7202..7223)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7463..7482)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    7644..7778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7668..7687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7722..7741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7921..7943)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 accccggggt ccagtgcttg ggccttctcc tgggtcatct gcggggccct gctctatcgc
       61 tcccgggggc acgtcaggct caccatctgg gccaccttct tggtggtatt caaaataatc
      121 ggcttcccct acagggtgga aaaatggcct tctacctgga gggggcctgc gcggtggaga
      181 cccggatgat gatgactgac tactgggact cctgggcctc ttttctccac gtccacgacc
      241 tctccccctg gctctttcac gacttccccc cctggctctt tcacgtcctc taccccggcg
      301 gcctccacta cctcctcgac cccggcctcc actacctcct cgaccccggc ctccactgcc
      361 tcctcgaccc cggcctccac ctcctgctcc tgcccctcct gctcctgccc ctcctcctgc
      421 tcctgcccct cctcctgnnc tcctgcccct cctgctcctg cccctcccgc tcctgctcct
      481 gctcctgttc caccgtgggt ccctttgcag ccaatgcaac ttggacgttt ttggggtctc
      541 cggacaccat ctctatgtct tggccctgat cctgagccgc ccggggctcc tggtcttccg
      601 cctcctcgtc ctcgtcctct tccccgtcct cgtccatggt tatcaccccc tcttctttga
      661 ggtccactgc cgccggagcc ttctggtcca gatgtgtctc ccttctctcc taggccattt
      721 ccaggtcctg tacctggccc ctcgtcagac atgattcaca ctaaaagaga tcaatagaca
      781 tctttattag acgacgctca gtgaatacag ggagtgcaga ctcctgcccc ctccaacagc
      841 ccccccaccc tcatcccctt catggtcgct gtcagacaga tccaggtctg aaaattcccc
      901 atcctccgaa ccatcctcgt cctcatcacc aattactcgc agcccggaaa actcccgctg
      961 aacatcctca agatttgcgt cctgagcctc aagccaggcc tcaaattcct cgtccccctt
     1021 tttgctggac ggtagggatg gggattctcg ggacccctcc tcttcctctt caaggtcacc
     1081 agacagagat gctactgggg caacggaaga aaagctgggt gcggcctgtg aggatcagct
     1141 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     1201 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1621 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1921 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2221 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2521 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttgaag ctgtccctga
     3061 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     3121 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     3181 acgtagccca gcgcgtcggc cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     3241 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     3301 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccctgct tcatccccgt
     3361 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
     3421 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
     3481 gcagtcgggg cggcgcggtc cgaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
     3541 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
     3601 ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     3661 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     3721 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     3781 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     3841 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     3901 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     3961 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     4021 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     4081 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     4141 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     4201 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     4261 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     4321 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     4381 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     4441 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     4501 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     4561 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     4621 cgtccggatc gggagatggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4681 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     4741 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4801 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4861 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc actggccccg
     4921 tgggttaggg acggggtccc ccatggggaa tggtttatgg ttcgtggggg ttattatttt
     4981 gggcgttgcg tggggtcagg tccacgactg gactgagcag acagacccat ggtttttgga
     5041 tggcctgggc atggaccgca tgtactggcg cgacacgaac accgggcgtc tgtggctgcc
     5101 aaacaccccc gacccccaaa aaccaccgcg cggatttctg gcgtgccaag ctagtcgacc
     5161 aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca
     5221 ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat
     5281 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     5341 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca
     5401 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5461 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5521 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5581 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5641 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5701 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     5761 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     5821 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5881 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     5941 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     6001 ctctagaagc tgggtaccag ctgctagcgc caccatgaag accatcatcg ccctgagcta
     6061 catcttctgc ctggtgttcg ccgagcagaa attgatcagc gaggaggacc tgggcagccc
     6121 tggaggcagt tcaggaggcg gatctcactc catgaggtat ttcttcacat ccgtgtcccg
     6181 gcccggccgc ggggagcccc gcttcatcgc agtgggctac gtggacgaca cgcagttcgt
     6241 gcggttcgac agcgacgccg cgagccagag gatggagccg cgggcgccgt ggatagagca
     6301 ggagggtccg gagtattggg acggggagac acggaaacct aaggcccact cacagactca
     6361 ccgagtggac ctggggaccc tgcgcggcta ctacaaccag agcgaggccg gttctcacac
     6421 cgtccagagg atgtatggct gcgacgtggg gtcggactgg cgcttcctcc gcgggtacca
     6481 ccagtacgcc tacgacggca aggattacat cgccctgaaa gaggacctgc gctcttggac
     6541 cgcggcggac atggcagctc agaccaccaa gcacaagtgg gaggcggccc atgtggcgga
     6601 gcagttgaga gcctacctgg agggcacgtg cgtggagtgg ctccgcagat acctggagaa
     6661 cgggaaggag acgctgcagc gtacggacgc ccccaaaacg catatgactc accacgctgt
     6721 ctctgaccat gaagccaccc tgaggtgctg ggccctgagc ttctaccctg cggagatcac
     6781 actgacctgg cagcgggatg gggaggacca gacccaggac acggagctcg tggagaccag
     6841 gcctgcaggg gatggaacct tccagaagtg ggcggctgtg gtggtgcctt ctggacagga
     6901 gcagagatac acctgccatg tgcagcatga gggtttgccc aagcccctca ccctgagatg
     6961 ggagccgtct tcccagccca ccatccccat cgtgggcatc attgctggcc tggttctctt
     7021 tggagctgtg atcactggag ctgtggtcgc tgctgtgatg tggaggagga agagctcaga
     7081 tagaaaagga gggagctact ctcaggctgc aagcagtgac agtgcccagg gctctgatgt
     7141 gtctctcaca gcttgtaaag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     7201 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     7261 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     7321 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta
     7381 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     7441 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     7501 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     7561 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atctgaccgc
     7621 ctaactcgag gccggcaagg ccggatccag acatgataag atacattgat gagtttggac
     7681 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     7741 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     7801 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     7861 aatgtggtat ggctgattat gatccggctg cctcgcgcgt ttcggtgatg acggtgaaaa
     7921 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7981 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgag
     8041 gtcgactcta gaggatcgat ccccgccccg gacgaactaa acctgactac gacatctctg
     8101 ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa cttgccaact
     8161 gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc tctgactgta
     8221 gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt tcatcctgga
     8281 gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa ctcttggctg
     8341 aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa gactacggga
     8401 ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga ccctcaagag
     8461 ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta gcatatgctt
     8521 cccgggtagt agtatatact atccagacta accctaattc aatagcatat gttacccaac
     8581 gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca gcgatatctc
     8641 ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg agggtagtga
     8701 accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga actctcctga
     8761 atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc tgagttgtga
     8821 acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa
     8881 atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac cctcacaaac
     8941 cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt aacaatagaa
     9001 atccatg
//
