LOCUS       sequence-335671-        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of huamn PD-L1 wildtype 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 WT
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1157..1180
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1225..1436
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1449..1717
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1744..2343
                     /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(1744..1763)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2240..2260
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2350..2938
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2403..2423)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2821..2832)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3012..3245
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3317..3438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3354..3373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3408..3427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3478..3613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3646..3662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3659..3681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3670..3686
                     /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(3694..3700,3701..3718,3719..3724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3877..3894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4807..5598,5599..5667))
                     /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     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5840..5858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5896..5918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6018..6037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6231..6253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6245..6262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6246..6262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6277..6503
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6504..6684
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6731..6856
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7349..7582
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7767..7811
                     /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    8078..8194
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggttt atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagattc tagagctagc gaattcatga ggatatttgc
      301 tgtctttata ttcatgacct actggcattt gctgaacgca tttactgtca cggttcccaa
      361 ggacctatat gtggtagagt atggtagcaa tatgacaatt gaatgcaaat tcccagtaga
      421 aaaacaatta gacctggctg cactaattgt ctattgggaa atggaggata agaacattat
      481 tcaatttgtg catggagagg aagacctgaa ggttcagcat agtagctaca gacagagggc
      541 ccggctgttg aaggaccagc tctccctggg aaatgctgca cttcagatca cagatgtgaa
      601 attgcaggat gcaggggtgt accgctgcat gatcagctat ggtggtgccg actacaagcg
      661 aattactgtg aaagtcaatg ccccatacaa caaaatcaac caaagaattt tggttgtgga
      721 tccagtcacc tctgaacatg aactgacatg tcaggctgag ggctacccca aggccgaagt
      781 catctggaca agcagtgacc atcaagtcct gagtggtaag accaccacca ccaattccaa
      841 gagagaggag aagcttttca atgtgaccag cacactgaga atcaacacaa caactaatga
      901 gattttctac tgcactttta ggagattaga tcctgaggaa aaccatacag ctgaattggt
      961 catcccagaa ctacctctgg cacatcctcc aaatgaaagg actcacttgg taattctggg
     1021 agccatctta ttatgccttg gtgtagcact gacattcatc ttccgtttaa gaaaagggag
     1081 aatgatggat gtgaaaaaat gtggcatcca agatacaaac tcaaagaagc aaagtgatac
     1141 acatttggag gagacggact acaaagacga tgacgacaag tgagcggccg cgaaggatct
     1201 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     1261 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     1321 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     1381 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     1441 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     1501 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     1561 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     1621 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     1681 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc
     1741 tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     1801 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     1861 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     1921 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     1981 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     2041 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     2101 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     2161 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     2221 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     2281 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     2341 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2761 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2941 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     3001 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg
     3061 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3121 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3181 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     3241 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     3301 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3361 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3421 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     3481 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3541 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3601 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5881 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5941 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6001 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6061 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6241 acgttgtaaa acgacggcca gtgccaagct gacgcgtgta gtcttatgca atactcttgt
     6301 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     6361 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     6421 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     6481 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     6541 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6601 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6661 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     6721 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     6781 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     6841 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     6901 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     6961 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     7021 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     7081 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     7141 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccac
     7201 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     7261 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     7321 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     7381 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     7441 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     7501 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     7561 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     7621 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     7681 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     7741 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     7801 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     7861 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     7921 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     7981 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     8041 ttcgattagt gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg
     8101 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     8161 aattacaaaa acaaattaca aaattcaaaa ttttatcgat actagtatta tgcccagtac
     8221 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8281 atg
//
