LOCUS       Exported                8425 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expression of HLA-DRA*01:01 cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    HLA_DRA_01_01
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8425)
  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 8425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8425
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1432..2020
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1485..1505)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1903..1914)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2092..2325
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2403..2524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2440..2459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2494..2513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2564..2699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2626..2645
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2719..2738)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2720..2738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2748..2765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2748..2764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2757..2779)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2906..3361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2993..3012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3203..3224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3387..3491
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3492..4352
                     /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(3710..3729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4523..5111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5012..5031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5265..5282
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5399..5420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5435..5465
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5473..5489
                     /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     5478..5500
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5497..5513
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5497..5513
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5532..5552
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5534..5552
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5580..5806
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5807..5987
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6034..6159
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6652..6885
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7070..7114
                     /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        7289..7789
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7708..7726
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7801..8196
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8256..8373
                     /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 catggccata agtggagtcc ctgtgctagg atttttcatc atagctgtgc
      661 tgatgagcgc tcaggaatca tgggctatca aagaagaaca tgtgatcatc caggccgagt
      721 tctatctgaa tcctgaccaa tcaggcgagt ttatgtttga ctttgatggt gatgagattt
      781 tccatgtgga tatggcaaag aaggagacgg tctggcggct tgaagaattt ggacgatttg
      841 ccagctttga ggctcaaggt gcattggcca acatagctgt ggacaaagcc aacctggaaa
      901 tcatgacaaa gcgctccaac tatactccga tcaccaatgt acctccagag gtaactgtgc
      961 tcacgaacag ccctgtggaa ctgagagagc ccaacgtcct catctgtttc atcgacaagt
     1021 tcaccccacc agtggtcaat gtcacgtggc ttcgaaatgg aaaacctgtc accacaggag
     1081 tgtcagagac agtcttcctg cccagggaag accacctttt ccgcaagttc cactatctcc
     1141 ccttcctgcc ctcaactgag gacgtttacg actgcagggt ggagcactgg ggcttggatg
     1201 agcctcttct caagcactgg gagtttgatg ctccaagccc tctcccagag actacagaga
     1261 acgtggtgtg tgccctgggc ctgactgtgg gtctggtggg catcattatt gggaccatct
     1321 tcatcatcaa gggagtgcgc aaaagcaatg cagcagaacg cagggggcct ctgtaatgta
     1381 caaagtggtt tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga caatcaacct
     1441 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1501 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1561 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1621 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1681 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1741 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1801 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     1861 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     1921 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     1981 cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga
     2041 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     2101 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     2161 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     2221 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     2281 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt
     2341 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     2401 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2461 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2521 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     2581 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2701 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     2761 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2821 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2881 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3301 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3361 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3421 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3481 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3601 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3661 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3721 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3781 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3901 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4021 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4081 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4321 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4381 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4441 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4501 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4561 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4621 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4681 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4741 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4801 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4861 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4921 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4981 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5041 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5101 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5161 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5221 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5281 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5341 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5401 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5461 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5521 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     5581 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     5641 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     5701 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     5761 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     5821 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5881 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5941 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     6001 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     6061 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     6121 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     6181 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     6241 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     6301 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     6361 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     6421 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     6481 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     6541 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     6601 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     6661 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     6721 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     6781 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     6841 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     6901 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     6961 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     7021 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     7081 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     7141 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     7201 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     7261 gacccacctc ccaaccccga ggggaccctt gcgccttttc caaggcagcc ctgggtttgc
     7321 gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc
     7381 tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg
     7441 ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc
     7501 gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg
     7561 ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc
     7621 agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt
     7681 agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac
     7741 gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg gggtaccacc
     7801 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     7861 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     7921 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     7981 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     8041 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     8101 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     8161 cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc agctgcagta gtaagaattc
     8221 tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     8281 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     8341 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     8401 ccactttggc gccggctcga ggggg
//
