LOCUS       sequence-327889-        8744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HLA-B*51:01 cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8744)
  AUTHORS   Lee MN, Meyerson M
  TITLE     Antigen identification for HLA class I- and HLA class II-restricted 
            T cell receptors using cytokine-capturing antigen-presenting cells.
  JOURNAL   Sci Immunol. 2021 Jan 22;6(55). pii: 6/55/eabf4001. doi: 
            10.1126/sciimmunol.abf4001.
  PUBMED    33483338
REFERENCE   2  (bases 1 to 8744)
  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"; journalName: "Sci 
            Immunol."; date: "2021-01-22"; volume: "6(55). pii: 6/55/eabf4001. 
            doi"; pages: " 10.1126/sciimmunol.abf4001"
COMMENT     Sequence Label: HLA_B5101
FEATURES             Location/Qualifiers
     source          1..8744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1751..2339
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1804..1824)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2222..2233)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2411..2644
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2722..2843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2759..2778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2813..2832
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2883..3018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3038..3057)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3039..3057)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3067..3084)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3067..3083)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3076..3098)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3225..3680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3312..3331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3522..3543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3706..3810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3811..3879,3880..4671)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4029..4048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4842..5430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5331..5350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5584..5601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5718..5739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5754..5759,5760..5777,5778..5784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5792..5808
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5797..5819
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5816..5832
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5816..5832
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5851..5871
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5853..5871
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5899..6125
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6126..6306
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6353..6478
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6971..7204
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7389..7433
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7608..8108
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8027..8045
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             8120..8515
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8575..8692
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgccgccac catgcgggtc acggcgcccc gaaccgtcct cctgctgctc tggggggcag
      661 tggccctgac cgagacctgg gccggctccc actccatgag gtatttctac accgccatgt
      721 cccggcccgg ccgcggggag ccccgcttca tcgcagtggg ctacgtggac gacacccagt
      781 tcgtgaggtt cgacagcgac gccgcgagtc cgaggacgga gccccgggcg ccatggatag
      841 agcaggaggg gccggagtat tgggaccgga acacacagat cttcaagacc aacacacaga
      901 cttaccgaga gaacctgcgg atcgcgctcc gctactacaa ccagagcgag gccgggtctc
      961 acacttggca gacgatgtat ggctgcgacg tggggccgga cgggcgcctc ctccgcgggc
     1021 ataaccagta cgcctacgac ggcaaagatt acatcgccct gaacgaggac ctgagctcct
     1081 ggaccgcggc ggacaccgcg gctcagatca cccagcgcaa gtgggaggcg gcccgtgagg
     1141 cggagcagct gagagcctac ctggagggcc tgtgcgtgga gtggctccgc agacacctgg
     1201 agaacgggaa ggagacgctg cagcgcgcgg accccccaaa gacacacgtg acccaccacc
     1261 ccgtctctga ccatgaggcc accctgaggt gctgggccct gggcttctac cctgcggaga
     1321 tcacactgac ctggcagcgg gatggcgagg accaaactca ggacactgag cttgtggaga
     1381 ccagaccagc aggagataga accttccaga agtgggcagc tgtggtggtg ccttctggag
     1441 aagagcagag atacacatgc catgtacagc atgaggggct gccgaagccc ctcaccctga
     1501 gatgggagcc atcttcccag tccaccatcc ccatcgtggg cattgttgct ggcctggctg
     1561 tcctagcagt tgtggtcatc ggagctgtgg tcgctactgt gatgtgtagg aggaagagct
     1621 caggtggaaa aggagggagc tactctcagg ctgcgtccag cgacagtgcc cagggctctg
     1681 atgtgtctct cacagcttga aaagtggttt agtaatgagc tagcgctaac cggtggcgcg
     1741 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1801 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1861 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1921 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1981 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2041 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2101 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2161 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2221 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2281 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     2341 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2401 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     2461 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     2521 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2581 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2641 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2701 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     2761 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2821 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     2881 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2941 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3001 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     3061 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3121 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3181 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3661 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4021 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4321 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5641 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5701 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5761 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5821 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     5881 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     5941 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     6001 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     6061 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     6121 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6181 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6241 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6301 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     6361 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     6421 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     6481 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     6541 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     6601 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     6661 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     6721 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     6781 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     6841 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     6901 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     6961 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     7021 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     7081 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     7141 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     7201 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     7261 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     7321 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     7381 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     7441 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     7501 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     7561 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccttg cgccttttcc
     7621 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     7681 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     7741 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     7801 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     7861 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     7921 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     7981 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     8041 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     8101 ctccccaggg ggtaccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     8161 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     8221 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     8281 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     8341 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     8401 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     8461 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggcctgca
     8521 gctgcagtag taagaattct agatcttgag acaaatggca gtattcatcc acaattttaa
     8581 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     8641 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     8701 ttacagggac agcagagatc cactttggcg ccggctcgag gggg
//
