LOCUS       sequence-236035-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-human PD-L1 in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  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: pGEX/PD-L1 WT
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             215..742
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(235..252)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(235..251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(244..266)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(844..849,850..865,866..872)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    880..896
                     /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)."
     CDS             919..1572
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1530..1552
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1579..1602
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2566..2588)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2733..2751)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2819..2923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2924..2992,2993..3784)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3142..3161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3955..4543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4444..4463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4697..4714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4787..4864
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4865..5855,1..92)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4884..4903)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg attcactggc
      241 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      301 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      361 ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc
      421 ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc
      481 aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac
      541 ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa
      601 tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttggaat
      661 tagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
      721 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
      781 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
      841 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
      901 acacaggaaa cagtattcat gtcccctata ctaggttatt ggaaaattaa gggccttgtg
      961 caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag
     1021 cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat
     1081 cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat
     1141 atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg
     1201 cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac
     1261 tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa
     1321 gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg
     1381 ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca
     1441 aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa
     1501 tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac
     1561 catcctccaa aatcggatct ggaagttctg ttccaggggc ccctgggatc cccggaattc
     1621 gcccttgtcg acatgaggat atttgctgtc tttatattca tgacctactg gcatttgctg
     1681 aacgcattta ctgtcacggt tcccaaggac ctatatgtgg tagagtatgg tagcaatatg
     1741 acaattgaat gcaaattccc agtagaaaaa caattagacc tggctgcact aattgtctat
     1801 tgggaaatgg aggataagaa cattattcaa tttgtgcatg gagaggaaga cctgaaggtt
     1861 cagcatagta gctacagaca gagggcccgg ctgttgaagg accagctctc cctgggaaat
     1921 gctgcacttc agatcacaga tgtgaaattg caggatgcag gggtgtaccg ctgcatgatc
     1981 agctatggtg gtgccgacta caagcgaatt actgtgaaag tcaatgcccc atacaacaaa
     2041 atcaaccaaa gaattttggt tgtggatcca gtcacctctg aacatgaact gacatgtcag
     2101 gctgagggct accccaaggc cgaagtcatc tggacaagca gtgaccatca agtcctgagt
     2161 ggtaagacca ccaccaccaa ttccaagaga gaggagaagc ttttcaatgt gaccagcaca
     2221 ctgagaatca acacaacaac taatgagatt ttctactgca cttttaggag attagatcct
     2281 gaggaaaacc atacagctga attggtcatc ccagaactac ctctggcaca tcctccaaat
     2341 gaaaggactc acttggtaat tctgggagcc atcttattat gccttggtgt agcactgaca
     2401 ttcatcttcc gtttaagaaa agggagaatg atggatgtga aaaaatgtgg catccaagat
     2461 acaaactcaa agaagcaaag tgatacacat ttggaggaga cgtaagcggc cgcatcgtga
     2521 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2581 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2641 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     2701 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     2761 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2821 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2881 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2941 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3001 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3061 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3121 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3181 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3241 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3301 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3361 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3421 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3481 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3541 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3601 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3661 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3721 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3781 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4081 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4621 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4681 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4741 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     4801 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     4861 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     4921 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     4981 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5041 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     5101 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     5161 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     5221 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     5281 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     5341 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     5401 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     5461 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     5521 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     5581 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     5641 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     5701 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     5761 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     5821 gcccgtctca ctggtgaaaa gaaaaaccac gctag
//
