LOCUS       sequence-270579-        9567 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged H2-Ld.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9567)
  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 9567)
  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-H2-Ld-VN
FEATURES             Location/Qualifiers
     source          1..9567
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(715..733)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        801..905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(906..974,975..1766)
                     /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(1124..1143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1937..2525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2426..2445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2956..3101
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3144..4181
                     /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(4179..4198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4223..4271
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4925..5304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5305..5508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5459..5479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5505..5529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5611..5640
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6679..6681,6682..7140)
                     /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(6721..6742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6982..7001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    7163..7297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7187..7206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7241..7260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7440..7462)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctgctcctgc tcctgttcca ccgtgggtcc ctttgcagcc aatgcaactt ggacgttttt
       61 ggggtctccg gacaccatct ctatgtcttg gccctgatcc tgagccgccc ggggctcctg
      121 gtcttccgcc tcctcgtcct cgtcctcttc cccgtcctcg tccatggtta tcaccccctc
      181 ttctttgagg tccactgccg ccggagcctt ctggtccaga tgtgtctccc ttctctccta
      241 ggccatttcc aggtcctgta cctggcccct cgtcagacat gattcacact aaaagagatc
      301 aatagacatc tttattagac gacgctcagt gaatacaggg agtgcagact cctgccccct
      361 ccaacagccc ccccaccctc atccccttca tggtcgctgt cagacagatc caggtctgaa
      421 aattccccat cctccgaacc atcctcgtcc tcatcaccaa ttactcgcag cccggaaaac
      481 tcccgctgaa catcctcaag atttgcgtcc tgagcctcaa gccaggcctc aaattcctcg
      541 tccccctttt tgctggacgg tagggatggg gattctcggg acccctcctc ttcctcttca
      601 aggtcaccag acagagatgc tactggggca acggaagaaa agctgggtgc ggcctgtgag
      661 gatcagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
      721 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      901 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      961 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1021 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1081 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1141 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1201 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1261 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1321 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1381 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1441 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1501 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1561 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1621 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1681 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1741 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1801 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     2101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttgaagct
     2581 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     2641 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     2701 ccagcaagac gtagcccagc gcgtcggccc cgagatgcgc cgcgtgcggc tgctggagat
     2761 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     2821 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccctgcttc
     2881 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     2941 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     3001 acgcagatgc agtcggggcg gcgcggtccg aggtccactt cgcatattaa ggtgacgcgt
     3061 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     3121 cagatcccgg ggggcaatga gatatgaaaa agcctgaact caccgcgacg tctgtcgaga
     3181 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     3241 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     3301 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     3361 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     3421 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     3481 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     3541 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     3601 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     3661 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     3721 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     3781 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     3841 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     3901 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     3961 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     4021 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     4081 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     4141 ccagcactcg tccggatcgg gagatggggg aggctaactg aaacacggaa ggagacaata
     4201 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     4261 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4321 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4381 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagccac
     4441 tggccccgtg ggttagggac ggggtccccc atggggaatg gtttatggtt cgtgggggtt
     4501 attattttgg gcgttgcgtg gggtcaggtc cacgactgga ctgagcagac agacccatgg
     4561 tttttggatg gcctgggcat ggaccgcatg tactggcgcg acacgaacac cgggcgtctg
     4621 tggctgccaa acacccccga cccccaaaaa ccaccgcgcg gatttctggc gtgccaagct
     4681 agtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca
     4741 ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa
     4801 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     4861 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     4921 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     4981 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     5041 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     5101 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5161 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     5221 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     5281 gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga
     5341 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     5401 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
     5461 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     5521 cgtcagatct ctagaagctg ggtaccagct gctagcgcca ccatgaagac catcatcgcc
     5581 ctgagctaca tcttctgcct ggtgttcgcc gagcagaaat tgatcagcga ggaggacctg
     5641 ggcagccctg gaggcagttc aggaggcgga ccacattcta tgcgttattt cgagaccgcc
     5701 gtgtcccggc ccggcctcgg ggagccccgg tacatctctg tcggctatgt ggacaacaag
     5761 gagttcgtgc gcttcgacag cgacgcggag aatccgagat atgagccgca ggcgccgtgg
     5821 atggagcagg aggggccgga gtattgggag cggatcacgc agatcgccaa gggccaggag
     5881 cagtggttcc gagtgaacct gaggaccctg ctcggctact acaaccagag cgcgggcggc
     5941 actcacacac tccagtggat gtacggctgt gacgtggggt cggacgggcg cctcctccgc
     6001 gggtacgagc agttcgccta cgacggctgc gattacatcg ccctgaacga agacctgaaa
     6061 acgtggacgg cggcggacat ggcggcgcag atcacccgac gcaagtggga gcaggctggt
     6121 gctgcagagt attacagggc ctacctggag ggcgagtgcg tggagtggct ccacagatac
     6181 ctgaagaacg ggaacgcgac gctgctgcgc acagattccc caaaggcaca tgtgacccat
     6241 caccccagat ctaaaggtga agtcaccctg aggtgctggg ccctgggctt ctaccctgct
     6301 gacatcaccc tgacctggca gttgaatggg gaggagctga cccaggacat ggagcttgtg
     6361 gagaccaggc ctgcagggga tggaaccttc cagaagtggg catctgtggt ggtgcctctt
     6421 gggaaggagc agaattacac atgccgtgtg taccatgagg ggctgcctga gcccctcacc
     6481 ctgagatggg agcctcctcc atctacggac agctacatgg tcatagtagc tgttcttggt
     6541 gtgctcgggg caatggcaat cattggtgca gtggtagcgt ttgtgatgaa acgccgacga
     6601 aataccggag gtaaaggtgg cgattatgcc ttggcaccag ggagccagtc atcagagatg
     6661 tcactgcgcg actgcaaggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     6721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     6781 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     6841 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     6901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     6961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tctgaccgcc
     7141 taactcgagg ccggcaaggc cggatccaga catgataaga tacattgatg agtttggaca
     7201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     7261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     7321 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     7381 atgtggtatg gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     7441 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     7501 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgagg
     7561 tcgactctag aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc
     7621 cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg
     7681 ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag
     7741 ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag
     7801 cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga
     7861 agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag
     7921 gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg
     7981 gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc
     8041 ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg
     8101 ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc
     8161 caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa
     8221 ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa
     8281 tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa
     8341 cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa
     8401 tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc
     8461 ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa
     8521 tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta
     8581 tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca
     8641 gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg
     8701 gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc
     8761 tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg
     8821 agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg
     8881 tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa
     8941 aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg
     9001 cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag
     9061 ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg
     9121 gtagcatata ctacccaaat atctggatag catatgctat cctaatctat atctgggtag
     9181 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     9241 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     9301 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     9361 catatgctat cctaatagag attagggtag tatatgctat cctaatttat atctgggtag
     9421 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     9481 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     9541 tgctatccta atctatatct gggtagt
//
