LOCUS       sequence-356707-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST fused human PD-1 intracellular domain (AA194-288).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  AUTHORS   Xu X, Hou B, Fulzele A, Masubuchi T, Zhao Y, Wu Z, Hu Y, Jiang Y, Ma
            Y, Wang H, Bennett EJ, Fu G, Hui E
  TITLE     PD-1 and BTLA regulate T cell signaling differentially and only 
            partially through SHP1 and SHP2.
  JOURNAL   J Cell Biol. 2020 Jun 1;219(6). pii: 151801. doi: 
            10.1083/jcb.201905085.
  PUBMED    32437509
REFERENCE   2  (bases 1 to 5264)
  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: "J Cell 
            Biol."; date: "2020-06-1"; volume: "219(6). pii: 151801. doi"; 
            pages: " 10.1083/jcb.201905085"
COMMENT     Sequence Label: pGEX-6p-2-GST-PD-1icd (AA194-288)
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(553..558,559..576,577..583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    591..607
                     /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     615..631
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     615..631
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             627..1154
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(647..664)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(647..663)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(656..678)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1256..1261,1262..1277,1278..1284)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1292..1308
                     /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             1331..1984
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1942..1964
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1991..2014
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2387..2409)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2554..2572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2640..2744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2745..2813,2814..3605)
                     /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(2963..2982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3776..4364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4265..4284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4518..4535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4608..4685
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4686..5264,1..504)
                     /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(4705..4724)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
       61 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
      121 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
      181 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
      241 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
      301 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
      361 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccacgct agcgcccaat
      421 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      481 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
      541 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      601 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
      661 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      721 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      781 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
      841 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
      901 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
      961 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     1021 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     1081 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     1141 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     1201 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     1261 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
     1321 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1381 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1441 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1501 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     1561 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     1621 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     1681 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     1741 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     1801 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     1861 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     1921 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     1981 aaaatcggat ctggaagttc tgttccaggg gcccctggga tcccgggccg cacgagggac
     2041 aataggagcc aggcgcaccg gccagcccct gaaggaggac ccctcagccg tgcctgtgtt
     2101 ctctgtggac tatggggagc tggatttcca gtggcgagag aagaccccgg agccccccgt
     2161 gccctgtgtc cctgagcaga cggagtatgc caccattgtc tttcctagcg gaatgggcac
     2221 ctcatccccc gcccgcaggg gctcagccga cggccctcgg agtgcccagc cactgaggcc
     2281 tgaggatgga cactgctctt ggcccctgta acccgggtcg actcgagcgg ccgcatcgtg
     2341 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2401 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2461 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2521 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     2581 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2641 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2701 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2761 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2821 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2881 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2941 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     3001 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3061 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3121 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3181 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3241 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     3301 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3361 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3421 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3481 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3541 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3601 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3661 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3901 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4021 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4141 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4381 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4441 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4501 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4561 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     4621 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     4681 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     4741 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     4801 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     4861 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     4921 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     4981 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5041 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     5101 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     5161 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     5221 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcg
//
