LOCUS       sequence-270555-        8310 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Mammalian expression of VN-fused and myc-tagged H2-Dd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8310)
  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 8310)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Dd-VN
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(55..73)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        141..245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(246..314,315..1106)
                     /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(464..483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1277..1865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1766..1785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2296..2441
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2484..3521
                     /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(3519..3538)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3563..3611
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4265..4644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4645..4848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4799..4819
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4845..4869
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4951..4980
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6028..6030,6031..6489)
                     /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(6070..6091)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6331..6350)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    6512..6646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6536..6555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6590..6609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6789..6811)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gatcagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
       61 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      481 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      781 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttgaagct
     1921 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     1981 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     2041 ccagcaagac gtagcccagc gcgtcggccc cgagatgcgc cgcgtgcggc tgctggagat
     2101 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     2161 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccctgcttc
     2221 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     2281 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     2341 acgcagatgc agtcggggcg gcgcggtccg aggtccactt cgcatattaa ggtgacgcgt
     2401 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     2461 cagatcccgg ggggcaatga gatatgaaaa agcctgaact caccgcgacg tctgtcgaga
     2521 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     2581 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     2641 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     2701 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     2761 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     2821 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     2881 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     2941 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     3001 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     3061 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     3121 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     3181 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     3241 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     3301 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     3361 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     3421 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     3481 ccagcactcg tccggatcgg gagatggggg aggctaactg aaacacggaa ggagacaata
     3541 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3601 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3661 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3721 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagccac
     3781 tggccccgtg ggttagggac ggggtccccc atggggaatg gtttatggtt cgtgggggtt
     3841 attattttgg gcgttgcgtg gggtcaggtc cacgactgga ctgagcagac agacccatgg
     3901 tttttggatg gcctgggcat ggaccgcatg tactggcgcg acacgaacac cgggcgtctg
     3961 tggctgccaa acacccccga cccccaaaaa ccaccgcgcg gatttctggc gtgccaagct
     4021 agtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca
     4081 ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa
     4141 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     4201 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     4261 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     4321 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4381 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4441 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4501 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     4561 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     4621 gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga
     4681 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4741 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
     4801 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     4861 cgtcagatct ctagaagctg ggtaccagct gctagcgcca ccatgaagac catcatcgcc
     4921 ctgagctaca tcttctgcct ggtgttcgcc gagcagaaat tgatcagcga ggaggacctg
     4981 ggcagccctg gaggcagttc aggaggcgga tcacactcgc tgaggtattt cgtcaccgcc
     5041 gtgtcccggc ccggcttcgg ggagccccgg tacatggaag tcggctacgt ggacaacacg
     5101 gagttcgtgc gcttcgacag cgacgcggag aatccgagat atgagccgcg ggcgcggtgg
     5161 atagagcagg aggggccgga gtattgggag cgggagacac gaagagccaa gggcaatgag
     5221 cagagtttcc gagtggacct gaggaccgcg ctgcgctact acaaccagag cgcgggcggc
     5281 tctcacacac tccagtggat ggctggctgt gacgtggagt cggacgggcg cctcctccgc
     5341 gggtactggc agttcgccta cgacggcagc gattacatcg ccctgaacga agacctgaaa
     5401 acgtggacgg cggcggacat ggcggcgcag atcacccgac gcaagtggga gcaggctggt
     5461 gctgcagaga gagaccgggc ctacctggag ggcgagtgcg tggagtggct ccgcagatac
     5521 ctgaagaacg ggaatgctac gctgctgcgc acagatcccc caaaggccca tgtgacccat
     5581 caccgcagac ctgaaggtga tgtcaccctg aggtgctggg ccctgggctt ctaccctgct
     5641 gacatcaccc tgacctggca gttgaatggg gaggagctga cccaggaaat ggagcttgtg
     5701 gagaccaggc ctgcagggga tggaaccttc cagaagtggg catctgtggt ggtgcctctt
     5761 gggaaggagc agaagtacac atgccatgtg gaacatgagg ggctgcctga gcccctcacc
     5821 ctgagatggg gcaaggagga acccccatcc tcaacaaaga ctaatactgt tattatcgca
     5881 gtaccagtag ttttgggtgc agtagtaatt ttgggagccg taatggcatt tgttatgaag
     5941 aggcggcgca acactggtgg gaagggcgga gactatgctt tggcacccgg cagccagtcc
     6001 tctgacatgt ccttgcctga ctgtaaggtg agcaagggcg aggagctgtt caccggggtg
     6061 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6121 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     6181 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     6241 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6301 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6361 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6421 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     6481 ctgaccgcct aactcgaggc cggcaaggcc ggatccagac atgataagat acattgatga
     6541 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6601 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     6661 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     6721 cctctacaaa tgtggtatgg ctgattatga tccggctgcc tcgcgcgttt cggtgatgac
     6781 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     6841 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     6901 gccatgaggt cgactctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     6961 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     7021 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     7081 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     7141 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     7201 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     7261 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     7321 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     7381 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     7441 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     7501 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     7561 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     7621 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     7681 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     7741 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     7801 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     7861 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     7921 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     7981 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     8041 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     8101 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     8161 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     8221 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     8281 tgcccacaaa accactaatg gcaccccggg
//
