LOCUS       sequence-464295-        5581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses CD86 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5581)
  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 5581)
  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-CD86
FEATURES             Location/Qualifiers
     source          1..5581
                     /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             2323..2352
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2673..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2710..2729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2764..2783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2822..2839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2993..3581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3073..3092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3752..4543,4544..4612))
                     /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     4375..4394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4613..4717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4785..4803
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4841..4863)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4963..4982
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5049..5504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5186..5207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5398..5417
                     /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 cgcggatcca tggaccccca gtgcactatg ggactgagta acattctctt tgtgatggcc
     1381 ttcctgctct ctggtgctgc tcctctgaag attcaagctt atttcaatga gactgcagac
     1441 ctgccatgcc aatttgcaaa ctctcaaaac caaagcctga gtgagctagt agtattttgg
     1501 caggaccagg aaaacttggt tctgaatgag gtatacttag gcaaagagaa atttgacagt
     1561 gttcattcca agtatatggg ccgcacaagt tttgattcgg acagttggac cctgagactt
     1621 cacaatcttc agatcaagga caagggcttg tatcaatgta tcatccatca caaaaagccc
     1681 acaggaatga ttcgcatcca ccagatgaat tctgaactgt cagtgcttgc taacttcagt
     1741 caacctgaaa tagtaccaat ttctaatata acagaaaatg tgtacataaa tttgacctgc
     1801 tcatctatac acggttaccc agaacctaag aagatgagtg ttttgctaag aaccaagaat
     1861 tcaactatcg agtatgatgg tattatgcag aaatctcaag ataatgtcac agaactgtac
     1921 gacgtttcca tcagcttgtc tgtttcattc cctgatgtta cgagcaatat gaccatcttc
     1981 tgtattctgg aaactgacaa gacgcggctt ttatcttcac ctttctctat agagcttgag
     2041 gaccctcagc ctcccccaga ccacattcct tggattacag ctgtacttcc aacagttatt
     2101 atatgtgtga tggttttctg tctaattcta tggaaatgga agaagaagaa gcggcctcgc
     2161 aactcttata aatgtggaac caacacaatg gagagggaag agagtgaaca gaccaagaaa
     2221 agagaaaaaa tccatatacc tgaaagatct gatgaagccc agcgtgtttt taaaagttcg
     2281 aagacatctt catgcgacaa aagtgataca tgttttctcg agcaccatca ccatcaccat
     2341 caccatcacc attagtacca ggtaagtgta cccaattcgc cctatagtga gtcgtattac
     2401 aattcactcg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gagatccaat
     2461 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattcaca
     2521 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     2581 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2641 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2761 gttgtggttt gtccaaactc atcaatgtat cttaacgcgc gactctagac tcttccgctt
     2821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4741 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4801 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4861 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4921 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4981 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5041 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5101 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     5161 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     5221 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5281 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5341 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5401 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5461 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     5521 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctagag
     5581 t
//
