LOCUS       sequence-271148-        7177 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged HLA-A*03:01.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7177)
  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 7177)
  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-A3-VN
FEATURES             Location/Qualifiers
     source          1..7177
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(306..324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        392..496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(497..565,566..1357)
                     /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(715..734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1528..2116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2017..2036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2547..2692
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2735..3772
                     /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(3770..3789)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3814..3862
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4516..4895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4896..5099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5050..5070
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5096..5120
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5202..5231
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6279..6281,6282..6740)
                     /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(6321..6342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6582..6601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    6763..6897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6787..6806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6841..6860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7040..7062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccaggtctga aaattcccca tcctccgaac catcctcgtc ctcatcacca attactcgca
       61 gcccggaaaa ctcccgctga acatcctcaa gatttgcgtc ctgagcctca agccaggcct
      121 caaattcctc gtcccccttt ttgctggacg gtagggatgg ggattctcgg gacccctcct
      181 cttcctcttc aaggtcacca gacagagatg ctactggggc aacggaagaa aagctgggtg
      241 cggcctgtga ggatcagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg
      301 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
      361 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      421 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      481 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      541 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      601 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      661 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      721 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      781 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      841 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      901 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      961 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1021 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     1081 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1141 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1201 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1261 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1321 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1381 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1441 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1501 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1561 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1621 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1681 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1741 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1801 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1861 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1921 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1981 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2041 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2101 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2161 gccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     2221 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     2281 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc ccgagatgcg ccgcgtgcgg
     2341 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
     2401 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     2461 cgccctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     2521 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
     2581 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc gaggtccact tcgcatatta
     2641 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca
     2701 acagcgtgcc gcagatcccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac
     2761 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     2821 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     2881 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     2941 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     3001 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3061 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     3121 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     3181 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     3241 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     3301 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     3361 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     3421 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     3481 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     3541 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     3601 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     3661 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     3721 aaaccgacgc cccagcactc gtccggatcg ggagatgggg gaggctaact gaaacacgga
     3781 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3841 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3901 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3961 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4021 gccatagcca ctggccccgt gggttaggga cggggtcccc catggggaat ggtttatggt
     4081 tcgtgggggt tattattttg ggcgttgcgt ggggtcaggt ccacgactgg actgagcaga
     4141 cagacccatg gtttttggat ggcctgggca tggaccgcat gtactggcgc gacacgaaca
     4201 ccgggcgtct gtggctgcca aacacccccg acccccaaaa accaccgcgc ggatttctgg
     4261 cgtgccaagc tagtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     4321 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     4381 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     4441 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     4501 tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4561 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4621 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4681 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4741 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     4801 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     4861 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     4921 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4981 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     5041 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     5101 gtttagtgaa ccgtcagatc tctagaagct gggtaccagc tgctagcgcc accatgaaga
     5161 ccatcatcgc cctgagctac atcttctgcc tggtgttcgc cgagcagaaa ttgatcagcg
     5221 aggaggacct gggcagccct ggaggcagtt caggaggcgg atctcattct atgagatatt
     5281 tcttcacatc cgtgtcccgg cccggccgcg gggagccccg cttcatcgcc gtgggctacg
     5341 tggacgacac gcagttcgtg cggttcgaca gcgacgccgc gagccagagg atggagccgc
     5401 gggcgccgtg gatagagcag gaggggccgg agtattggga ccaggagaca cggaatgtga
     5461 aggcccagtc acagactgac cgagtggacc tggggaccct gcgcggctac tacaaccaga
     5521 gcgaggccgg ttctcacacc atccagataa tgtatggctg cgacgtgggg tcggacgggc
     5581 gcttcctccg cgggtaccgg caggacgcct acgacggcaa ggattacatc gccctgaacg
     5641 aggacctgcg ctcttggacc gcggcggaca tggcggctca gatcaccaag cgcaagtggg
     5701 aggcggccca tgaggcggag cagttgagag cctacctgga tggcacgtgc gtggagtggc
     5761 tccgcagata cctggagaac gggaaggaga cgctgcagcg cacggacccc cccaagacac
     5821 atatgaccca ccaccccatc tctgaccatg aggccaccct gaggtgctgg gccctgggct
     5881 tctaccctgc ggagatcaca ctgacctggc agcgggatgg ggaggaccag acccaggaca
     5941 cggagctcgt ggagaccagg cctgcagggg atggaacctt ccagaagtgg gcggctgtgg
     6001 tggtgccttc tggagaggag cagagataca cctgccatgt gcagcatgag ggtctgccca
     6061 agcccctcac cctgagatgg gagctgtctt cccagcccac tataccgatt gttggcatca
     6121 tagccggact tgtattgctg ggtgcggtga ttacgggcgc cgtcgtcgct gctgtgatgt
     6181 ggcggcggaa gtcttcagac agaaaaggcg gatcttatac ccaagctgca agctcagatt
     6241 cagcacaggg gtctgatgtg agtctgacag cttgcaaagt gagcaagggc gaggagctgt
     6301 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     6361 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     6421 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc
     6481 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     6541 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     6601 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     6661 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     6721 acaacgtcta tctgaccgcc taactcgagg ccggcaaggc cggatccaga catgataaga
     6781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     6901 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     6961 agcaagtaaa acctctacaa atgtggtatg gctgattatg atccggctgc ctcgcgcgtt
     7021 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     7081 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7141 gtcggggcgc agccatgagg tcgactctag aggatcg
//
