LOCUS       sequence-270553-        7171 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged HLA-A*02:01.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7171)
  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 7171)
  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
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(94..112)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        180..284
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(285..353,354..1145)
                     /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(503..522)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1316..1904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1805..1824
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2335..2480
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2523..3560
                     /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(3558..3577)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3602..3650
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4304..4683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4684..4887
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4838..4858
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4884..4908
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4990..5019
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6067..6069,6070..6528)
                     /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(6109..6130)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6370..6389)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    6551..6685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6575..6594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6629..6648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6828..6850)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 actggggcaa cggaagaaaa gctgggtgcg gcctgtgagg atcagcttat cgatgataag
       61 ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
      121 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
      181 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      241 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      301 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      361 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      421 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      481 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
      541 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
      601 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
      661 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
      721 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
      781 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
      841 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
      901 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
      961 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1021 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1081 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1141 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1201 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1261 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1321 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1381 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1441 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1501 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1561 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1621 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1681 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1741 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1801 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1861 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1921 cctttttacg gttcctggcc ttttgctggc cttgaagctg tccctgatgg tcgtcatcta
     1981 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     2041 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     2101 cgtcggcccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     2161 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     2221 tggagtggtg aatccgttag cgaggtgccg ccctgcttca tccccgtggc ccgttgctcg
     2281 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
     2341 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
     2401 cgcggtccga ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
     2461 ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccggg gggcaatgag
     2521 atatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     2581 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     2641 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     2701 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     2761 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     2821 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     2881 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     2941 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     3001 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     3061 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     3121 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     3181 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     3241 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     3301 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     3361 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     3421 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     3481 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccggatcggg
     3541 agatggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3601 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3661 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3721 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3781 cgcagccaac gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg
     3841 gggtccccca tggggaatgg tttatggttc gtgggggtta ttattttggg cgttgcgtgg
     3901 ggtcaggtcc acgactggac tgagcagaca gacccatggt ttttggatgg cctgggcatg
     3961 gaccgcatgt actggcgcga cacgaacacc gggcgtctgt ggctgccaaa cacccccgac
     4021 ccccaaaaac caccgcgcgg atttctggcg tgccaagcta gtcgaccaat tctcatgttt
     4081 gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg
     4141 taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
     4201 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     4261 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
     4321 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     4381 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4441 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4501 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4561 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4621 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4681 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
     4741 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4801 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta
     4861 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatctc tagaagctgg
     4921 gtaccagctg ctagcgccac catgaagacc atcatcgccc tgagctacat cttctgcctg
     4981 gtgttcgccg agcagaaatt gatcagcgag gaggacctgg gcagccctgg aggcagttca
     5041 ggaggcggat ctcactccat gaggtatttc ttcacatccg tgtcccggcc cggccgcggg
     5101 gagccccgct tcatcgcagt gggctacgtg gacgacacgc agttcgtgcg gttcgacagc
     5161 gacgccgcga gccagaggat ggagccgcgg gcgccgtgga tagagcagga gggtccggag
     5221 tattgggacg gggagacacg gaaagtgaag gcccactcac agactcaccg agtggacctg
     5281 gggaccctgc gcggctacta caaccagagc gaggccggtt ctcacaccgt ccagaggatg
     5341 tatggctgcg acgtggggtc ggactggcgc ttcctccgcg ggtaccacca gtacgcctac
     5401 gacggcaagg attacatcgc cctgaaagag gacctgcgct cttggaccgc ggcggacatg
     5461 gcagctcaga ccaccaagca caagtgggag gcggcccatg tggcggagca gttgagagcc
     5521 tacctggagg gcacgtgcgt ggagtggctc cgcagatacc tggagaacgg gaaggagacg
     5581 ctgcagcgta cggacgcccc caaaacgcat atgactcacc acgctgtctc tgaccatgaa
     5641 gccaccctga ggtgctgggc cctgagcttc taccctgcgg agatcacact gacctggcag
     5701 cgggatgggg aggaccagac ccaggacacg gagctcgtgg agaccaggcc tgcaggggat
     5761 ggaaccttcc agaagtgggc ggctgtggtg gtgccttctg gacaggagca gagatacacc
     5821 tgccatgtgc agcatgaggg tttgcccaag cccctcaccc tgagatggga gccgtcttcc
     5881 cagcccacca tccccatcgt gggcatcatt gctggcctgg ttctctttgg agctgtgatc
     5941 actggagctg tggtcgctgc tgtgatgtgg aggaggaaga gctcagatag aaaaggaggg
     6001 agctactctc aggctgcaag cagtgacagt gcccagggct ctgatgtgtc tctcacagct
     6061 tgtaaagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     6121 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     6181 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     6241 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     6301 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     6361 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     6421 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     6481 cacaagctgg agtacaacta caacagccac aacgtctatc tgaccgccta actcgaggcc
     6541 ggcaaggccg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6601 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6661 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6721 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     6781 tgattatgat ccggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6841 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6901 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaggtc gactctagag
     6961 gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg
     7021 gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca
     7081 catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta
     7141 taaatggatg tgcacatttg ccaacactga g
//
