LOCUS       sequence-464235-        5488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses PD-L1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5488)
  AUTHORS   Chou WC, Chen WT, Kuo CT, Chang YM, Lu YS, Li CW, Hung MC, Shen CY
  TITLE     Genetic insights into carbohydrate sulfotransferase 8 and its impact
            on the immunotherapy efficacy of cancer.
  JOURNAL   Cell Rep. 2024 Jan 23;43(1):113641. doi: 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30.
  PUBMED    38165805
REFERENCE   2  (bases 1 to 5488)
  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: "Cell Rep.";
            date: "2024-01-23"; volume: "43(1):113641. doi"; pages: " 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30"
COMMENT     Sequence Label: pXJ-His-CD274
FEATURES             Location/Qualifiers
     source          1..5488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     506..530
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          671..1243
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1251..1270
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2230..2259
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2288..2306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2580..2701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2617..2636)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2671..2690
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2729..2746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2900..3488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2980..2999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3659..4450,4451..4519))
                     /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     4282..4301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4520..4624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4692..4710
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4748..4770)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4870..4889
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4956..5411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5093..5114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5305..5324
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      541 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
      601 gggaacggtg cattggaacg gacccctagg cttttgcaaa aagctccgga tcgatcctga
      661 gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgtaaaatt
      721 catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt
      781 gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc
      841 attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca
      901 tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt actttctcta
      961 atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac agttttagag
     1021 aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg
     1081 tggaaatatt cttattggta gaaacaacta catcctggtc atcatcctgc ctttctcttt
     1141 atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg
     1201 cccctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1261 ggttattgtg ctgtctcatc attttggcaa agaattgtaa tacgactcac taagggcggc
     1321 cgcggatccc ccgggctgca ggaattcgcc accatgagga tatttgctgt ctttatattc
     1381 atgacctact ggcatttgct gaacgcattt actgtcacgg ttcccaagga cctatatgtg
     1441 gtagagtatg gtagcaatat gacaattgaa tgcaaattcc cagtagaaaa acaattagac
     1501 ctggctgcac taattgtcta ttgggaaatg gaggataaga acattattca atttgtgcat
     1561 ggagaggaag acctgaaggt tcagcatagt agctacagac agagggcccg gctgttgaag
     1621 gaccagctct ccctgggaaa tgctgcactt cagatcacag atgtgaaatt gcaggatgca
     1681 ggggtgtacc gctgcatgat cagctatggt ggtgccgact acaagcgaat tactgtgaaa
     1741 gtcaatgccc catacaacaa aatcaaccaa agaattttgg ttgtggatcc agtcacctct
     1801 gaacatgaac tgacatgtca ggctgagggc taccccaagg ccgaagtcat ctggacaagc
     1861 agtgaccatc aagtcctgag tggtaagacc accaccacca attccaagag agaggagaag
     1921 cttttcaatg tgaccagcac actgagaatc aacacaacaa ctaatgagat tttctactgc
     1981 acttttagga gattagatcc tgaggaaaac catacagctg aattggtcat cccagaacta
     2041 cctctggcac atcctccaaa tgaaaggact cacttggtaa ttctgggagc catcttatta
     2101 tgccttggtg tagcactgac attcatcttc cgtttaagaa aagggagaat gatggatgtg
     2161 aaaaaatgtg gcatccaaga tacaaactca aagaagcaaa gtgatacaca tttggaggag
     2221 acgctcgagc accatcacca tcaccatcac catcaccatt agtaccaggt aagtgtaccc
     2281 aattcgccct atagtgagtc gtattacaat tcactcgatc gcccttccca acagttgcgc
     2341 agcctgaatg gcgaatggag atccaatttt taagtgtata atgtgttaaa ctactgattc
     2401 taattgtttg tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc
     2461 acagtctgtt catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2521 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2581 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2641 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2701 aacgcgcgac tctagactct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3301 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3661 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3721 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3781 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3841 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3901 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3961 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4021 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4081 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4141 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4201 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4261 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4321 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4381 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4441 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4501 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4561 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4621 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4681 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4741 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4801 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4861 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     4921 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     4981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5401 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5461 gcctcttcgc tattacgcca gctagagt
//
