LOCUS       sequence-327885-        8461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HLA-DRB1*04:01 cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8461)
  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 8461)
  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_DRB1_04_01
FEATURES             Location/Qualifiers
     source          1..8461
                     /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"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1468..2056
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1521..1541)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1939..1950)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2128..2361
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2439..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2476..2495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2530..2549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2600..2735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2755..2774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2756..2774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2784..2801)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2784..2800)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2793..2815)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2942..3397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3029..3048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3239..3260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3423..3527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3528..3596,3597..4388)
                     /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(3746..3765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4559..5147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5048..5067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5301..5318
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5435..5456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5471..5476,5477..5494,5495..5501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5509..5525
                     /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     5514..5536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5533..5549
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5533..5549
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5568..5588
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5570..5588
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5616..5842
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5843..6023
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6070..6195
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6688..6921
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7106..7150
                     /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        7325..7825
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7744..7762
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7837..8232
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8292..8409
                     /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 catggtgtgt ctgaagttcc ctggaggctc ctgcatggca gctctgacag
      661 tgacactgat ggtgctgagc tccccactgg ctttggctgg ggacacccga ccacgtttct
      721 tggagcaggt taaacatgag tgtcatttct tcaacgggac ggagcgggtg cggttcctgg
      781 acagatactt ctatcaccaa gaggagtacg tgcgcttcga cagcgacgtg ggggagtacc
      841 gggcggtgac ggagctgggg cggcctgatg ccgagtactg gaacagccag aaggacctcc
      901 tggagcagaa gcgggccgcg gtggacacct actgcagaca caactacggg gttggtgaga
      961 gcttcacagt gcagcggcga gtctatcctg aggtgactgt gtatcctgca aagacccagc
     1021 ccctgcagca ccacaacctc ctggtctgct ctgtgaatgg tttctatcca ggcagcattg
     1081 aagtcaggtg gttccggaac ggccaggaag agaagactgg ggtggtgtcc acaggcctga
     1141 tccagaatgg agactggacc ttccagaccc tggtgatgct ggaaacagtt cctcggagtg
     1201 gagaggttta cacctgccaa gtggagcacc caagcctgac gagccctctc acagtggaat
     1261 ggagagcacg gtctgaatct gcacagagca agatgctgag tggagtcggg ggcttcgtgc
     1321 tgggcctgct cttccttggg gccgggctgt tcatctactt caggaatcag aaaggacact
     1381 ctggacttca gccaacagga ttcctgagct gatgtacaaa gtggtttagt aatgagctag
     1441 cgctaaccgg tggcgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     1501 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1561 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1621 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     1681 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     1741 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     1801 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     1861 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     1921 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     1981 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2041 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
     2101 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     2161 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     2221 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     2281 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     2341 cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt
     2401 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     2461 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2521 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     2581 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2641 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2701 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     2761 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2821 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2881 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2941 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3361 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     3421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4441 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4501 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4561 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4621 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4681 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4741 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4801 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4861 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4921 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4981 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5041 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5101 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5161 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5221 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5281 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5341 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5401 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     5461 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5521 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     5581 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
     5641 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
     5701 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
     5761 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
     5821 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
     5881 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5941 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6001 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     6061 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     6121 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     6181 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     6241 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     6301 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     6361 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     6421 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     6481 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     6541 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     6601 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     6661 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     6721 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     6781 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     6841 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     6901 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     6961 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     7021 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     7081 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     7141 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     7201 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     7261 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     7321 acccttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg
     7381 tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg
     7441 cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc
     7501 gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc
     7561 cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg
     7621 accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg
     7681 ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc
     7741 gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga
     7801 ccgaatcacc gacctctctc cccagggggt accaccatgg ccaagccttt gtctcaagaa
     7861 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     7921 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     7981 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     8041 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     8101 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     8161 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     8221 gtgtgggagg gcctgcagct gcagtagtaa gaattctaga tcttgagaca aatggcagta
     8281 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     8341 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     8401 caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg
     8461 g
//
