LOCUS       sequence-488310-        6014 bp    DNA     circular SYN 03-OCT-2025
DEFINITION  expresses TAPBPR-S104F-K211L-R270Q in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  AUTHORS   Sun Y, Papadaki GF, Devlin CA, Danon JN, Young MC, Winters TJ, 
            Burslem GM, Procko E, Sgourakis NG
  TITLE     Xeno interactions between MHC-I proteins and molecular chaperones 
            enable ligand exchange on a broad repertoire of HLA allotypes.
  JOURNAL   Sci Adv. 2023 Feb 24;9(8):eade7151. doi: 10.1126/sciadv.ade7151. 
            Epub 2023 Feb 24.
  PUBMED    36827371
REFERENCE   2  (bases 1 to 6014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2023-02-24"; volume: "9(8):eade7151. doi"; pages: " 
            10.1126/sciadv.ade7151. Epub 2023 Feb 24"
COMMENT     Sequence Label: TAPBPR S104F K211L R270Q
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..24
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     16..33
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     17..33
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        39..465
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     RBS             861..869
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1738..1761
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1768..1812
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1819..1836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1852..1893
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1903..1920
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1943..1960)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1962..2096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1986..2005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2040..2059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2218..2817)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2301..2321)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2798..2817
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(3431..3447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3431..3447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3444..3466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3455..3471
                     /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)."
     promoter        complement(join(3479..3485,3486..3503,3504..3509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3662..3679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4592..5383,5384..5452))
                     /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     5215..5234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5453..5557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5625..5643
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5681..5703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5803..5822
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcccagtcac gacgttgtaa aacgacggcc agtgaattaa ttcgttgcag gacaggatgt
       61 ggtgcccgat gtgactagct ctttgctgca ggccgtccta tcctctggtt ccgataagag
      121 acccagaact ccggcccccc accgcccacc gccaccccca tacatatgtg gtacgcaagt
      181 aagagtgcct gcgcatgccc catgtgcccc accaagagct ttgcatccca tacaagtccc
      241 caaagtggag aaccgaacca attcttcgcg ggcagaacaa aagcttctgc acacgtctcc
      301 actcgaattt ggagccggcc ggcgtgtgca aaagaggtga atcgaacgaa agacccgtgt
      361 gtaaagccgc gtttccaaaa tgtataaaac cgagagcatc tggccaatgt gcatcagttg
      421 tggtcagcag caaaatcaag tgaatcatct cagtgcaact aaagggggga tctagatcgg
      481 ggtacctact agtccagtgt ggtggaattc atgggtactc aggagggatg gtgcctgttg
      541 ctgtgcttgg ccttaagcgg agctgctgag accaaaccgc atcccgcgga gggtcagtgg
      601 cgcgcggtcg atgtggtgct cgactgcttc ctagtcaagg atggagccca ccgaggcgcc
      661 ctcgcatcca gtgaggatcg agctcgtgca tccttggtac tgaagcaagt gccggtactg
      721 gacgacggat cgctcgaaga cttcaccgac tttcagggcg gaacgctggc gcaggatgat
      781 ccaccaatta tattcgaagc tagtgtggat ctggtacaga tcccgcaggc cgaagctctg
      841 ctgcacgccg atagctccgg taaggaggtt acctgcgaga tcttccgcta ctttctgcag
      901 atgaccgaaa cgacggtgaa aaccgccgct tggttcatgg ccaatgtgca agtttctggc
      961 ggtggaccgt ccattagcct ggtcatgaaa acgccacggg tggccaagaa cgaagtgctg
     1021 tggcatccga cgttgaatct gcctctctcc ccccaaggaa ctgtccgcac tgccgttgag
     1081 ttccaggtga tgacacagac ccagagccta tcctttttac taggctcatc agcgagcttg
     1141 gattgcggct tcagcatggc cccagggctc gacctcatat cggtggagtg gcggctccag
     1201 cacctgggca gaggccagtt ggtctattcg tggacggccg gtcaaggtca agcggtgcgc
     1261 aagggcgcaa cattggaacc ggcgcaactt ggcatggcca gggatgccag cctaaccttg
     1321 cccggcctga ctattcagga cgagggcacc tatatttgtc aaatcacaac atctctctac
     1381 caggctcagc agattataca actgaacatc caggcgtcgc ccaaggtgag gctgtcgctg
     1441 gccaacgagg cccttcttcc cacactgatc tgtgatattg ccggctacta ccccttagac
     1501 gtcgttgtca cctggactcg cgaggagctg ggtggctcac ccgctcaggt tagcggggcg
     1561 agtttttcgt ctcttcgtca atccgttgca ggaacctact ccatctcgag ttcactaact
     1621 gcagaacctg gctccgcagg agccacctat acatgccagg tgacccacat ctctctggag
     1681 gaacctctgg gggcctcgac gcaggttgtg ccgccagagc gaagattgga aggtggcctg
     1741 gaggtgctct tccagggacc cgggggtgga ctgaacgata tttttgaagc ccaaaagatc
     1801 gagtggcacg agggcggcca tcaccaccac catcattaag ggcccttcga aggtaagcct
     1861 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat
     1921 tgagtttaaa cccgctgatc agcctcgact gtgccttcta agatccagac atgataagat
     1981 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2041 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttctag
     2101 agtcggtggg cctcgggggc gggtgcgggg tcggcggggc cgccccgggt ggcttcggtc
     2161 ggagccatgg ggtcgtgcgc tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca
     2221 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
     2281 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag
     2341 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
     2401 cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc
     2461 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
     2521 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
     2581 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
     2641 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
     2701 cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
     2761 ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatgga
     2821 aggtcgtctc cttgtgaggg gtcaggggcg tgggtcaggg gatggtggcg gcaccggtcg
     2881 tggcggccga cctgcaggca tgcaagctat cgaattcctg cagcccgggg gatctgttgt
     2941 aatttataat ttatatttcc tttcttaata aataaataaa tagtcaagtt tatgtttgag
     3001 ttttatgatt tatattttaa gttatttcaa ctgcaacacc agcaccacga cctacttaca
     3061 gcaaaaaacg tacaagaagg aaagaaggaa taaaaagagt ggtattctct tacaatatgt
     3121 tttatggcat aaaaggtgtg gccattcata tcaaatataa agtagtgttg tttaacgtta
     3181 cttttgtagg ttgaatagta tattccaaca gatgatgagg ggttcccaat cctaaaccca
     3241 tttgccgttc ccagaagcat gaaaccacca cgcaccggat cctctagaac aacaacaatt
     3301 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3361 acctctacaa atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg
     3421 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3481 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5581 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5641 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5701 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5761 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     5821 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5881 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5941 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6001 taacgccagg gttt
//
