LOCUS       sequence-249794-        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human PD-L1 mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8283)
  AUTHORS   Li CW, Lim SO, Xia W, Lee HH, Chan LC, Kuo CW, Khoo KH, Chang SS, 
            Cha JH, Kim T, Hsu JL, Wu Y, Hsu JM, Yamaguchi H, Ding Q, Wang Y, 
            Yao J, Lee CC, Wu HJ, Sahin AA, Allison JP, Yu D, Hortobagyi GN, 
            Hung MC
  TITLE     Glycosylation and stabilization of programmed death ligand-1 
            suppresses T-cell activity.
  JOURNAL   Nat Commun. 2016 Aug 30;7:12632. doi: 10.1038/ncomms12632.
  PUBMED    27572267
REFERENCE   2  (bases 1 to 8283)
  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: "Nat 
            Commun."; date: "2016-08-30"; volume: "7:12632. doi"; pages: " 
            10.1038/ncomms12632"
COMMENT     Sequence Label: pCDH-puro/Flag PD-L1 4NQ
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     195..211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        226..452
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             453..633
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    680..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1298..1531
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1716..1760
                     /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"
     misc_feature    2027..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2233..2436
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2387..2407
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2433..2457
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3389..3412
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3457..3668
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3681..3949
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3976..4575
                     /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(3976..3995)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4472..4492
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4582..5170
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4635..4655)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5053..5064)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5244..5477
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5549..5670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5586..5605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5640..5659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5710..5845
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5878..5894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5878..5894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5891..5913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5902..5918
                     /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(5926..5932,5933..5950,5951..5956))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5971..5992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6109..6126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6280..6868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7039..7830,7831..7899))
                     /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     7662..7681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7900..8004)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8072..8090
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8128..8150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8250..8269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg acgcgtgtag tcttatgcaa
      241 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
      301 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
      361 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
      421 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
      481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccact gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     1201 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     1261 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     1321 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     1381 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     1441 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     1501 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     1561 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     1621 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     1681 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     1741 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     1801 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     1861 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     1921 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     1981 acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aattcaaaat tttatcgata ctagtattat
     2161 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2221 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2281 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2341 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2401 aggcgtgtac ggtgggaggt ttatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2461 tggagacgcc atccacgctg ttttgacctc catagaagat tctagagcta gcgaattcat
     2521 gaggatattt gctgtcttta tattcatgac ctactggcat ttgctgaacg catttactgt
     2581 cacggttccc aaggacctat atgtggtaga gtatggtagc caaatgacaa ttgaatgcaa
     2641 attcccagta gaaaaacaat tagacctggc tgcactaatt gtctattggg aaatggagga
     2701 taagaacatt attcaatttg tgcatggaga ggaagacctg aaggttcagc atagtagcta
     2761 cagacagagg gcccggctgt tgaaggacca gctctccctg ggaaatgctg cacttcagat
     2821 cacagatgtg aaattgcagg atgcaggggt gtaccgctgc atgatcagct atggtggtgc
     2881 cgactacaag cgaattactg tgaaagtcaa tgccccatac aacaaaatca accaaagaat
     2941 tttggttgtg gatccagtca cctctgaaca tgaactgaca tgtcaggctg agggctaccc
     3001 caaggccgaa gtcatctgga caagcagtga ccatcaagtc ctgagtggta agaccaccac
     3061 caccaattcc aagagagagg agaagctttt ccaagtgacc agcacactga gaatccagac
     3121 aacaactaat gagattttct actgcacttt taggagatta gatcctgagg aacagcatac
     3181 agctgaattg gtcatcccag aactacctct ggcacatcct ccaaatgaaa ggactcactt
     3241 ggtaattctg ggagccatct tattatgcct tggtgtagca ctgacattca tcttccgttt
     3301 aagaaaaggg agaatgatgg atgtgaaaaa atgtggcatc caagatacaa actcaaagaa
     3361 gcaaagtgat acacatttgg aggagacgga ctacaaagac gatgacgaca agtgagcggc
     3421 cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     3481 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg
     3541 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     3601 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     3661 gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct
     3721 gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct
     3781 gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg
     3841 cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg
     3901 ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac
     3961 cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4021 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4081 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4141 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4201 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4261 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4321 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4381 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4441 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4501 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4561 aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4621 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4681 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4741 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4801 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4861 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4921 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4981 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5041 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5101 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5161 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5221 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     5281 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5341 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5401 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5461 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     5521 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5581 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5641 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5701 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5761 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5821 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     5881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5941 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6721 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8041 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8101 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8161 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8221 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8281 cca
//
