LOCUS       sequence-464239-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses CD276 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  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 6196)
  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-CD276
FEATURES             Location/Qualifiers
     source          1..6196
                     /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             2938..2967
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2996..3014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3288..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3325..3344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3379..3398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3437..3454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3608..4196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4367..5158,5159..5227))
                     /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     4990..5009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5228..5332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5400..5418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5456..5478)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5578..5597
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5664..6119)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5801..5822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6013..6032
                     /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 tgctgcgtcg gcggggcagc cctggcatgg gtgtgcatgt gggtgcagcc
     1381 ctgggagcac tgtggttctg cctcacagga gccctggagg tccaggtccc tgaagaccca
     1441 gtggtggcac tggtgggcac cgatgccacc ctgtgctgct ccttctcccc tgagcctggc
     1501 ttcagcctgg cacagctcaa cctcatctgg cagctgacag ataccaaaca gctggtgcac
     1561 agctttgctg agggccagga ccagggcagc gcctatgcca accgcacggc cctcttcccg
     1621 gacctgctgg cacagggcaa cgcatccctg aggctgcagc gcgtgcgtgt ggcggacgag
     1681 ggcagcttca cctgcttcgt gagcatccgg gatttcggca gcgctgccgt cagcctgcag
     1741 gtggccgctc cctactcgaa gcccagcatg accctggagc ccaacaagga cctgcggcca
     1801 ggggacacgg tgaccatcac gtgctccagc taccagggct accctgaggc tgaggtgttc
     1861 tggcaggatg ggcagggtgt gcccctgact ggcaacgtga ccacgtcgca gatggccaac
     1921 gagcagggct tgtttgatgt gcacagcatc ctgcgggtgg tgctgggtgc aaatggcacc
     1981 tacagctgcc tggtgcgcaa ccccgtgctg cagcaggatg cgcacagctc tgtcaccatc
     2041 acaccccaga gaagccccac aggagccgtg gaggtccagg tccctgagga cccggtggtg
     2101 gccctagtgg gcaccgatgc caccctgcgc tgctccttct cccccgagcc tggcttcagc
     2161 ctggcacagc tcaacctcat ctggcagctg acagacacca aacagctggt gcacagtttc
     2221 accgaaggcc gggaccaggg cagcgcctat gccaaccgca cggccctctt cccggacctg
     2281 ctggcacaag gcaatgcatc cctgaggctg cagcgcgtgc gtgtggcgga cgagggcagc
     2341 ttcacctgct tcgtgagcat ccgggatttc ggcagcgctg ccgtcagcct gcaggtggcc
     2401 gctccctact cgaagcccag catgaccctg gagcccaaca aggacctgcg gccaggggac
     2461 acggtgacca tcacgtgctc cagctaccgg ggctaccctg aggctgaggt gttctggcag
     2521 gatgggcagg gtgtgcccct gactggcaac gtgaccacgt cgcagatggc caacgagcag
     2581 ggcttgtttg atgtgcacag cgtcctgcgg gtggtgctgg gtgcgaatgg cacctacagc
     2641 tgcctggtgc gcaaccccgt gctgcagcag gatgcgcacg gctctgtcac catcacaggg
     2701 cagcctatga cattcccccc agaggccctg tgggtgaccg tggggctgtc tgtctgtctc
     2761 attgcactgc tggtggccct ggctttcgtg tgctggagaa agatcaaaca gagctgtgag
     2821 gaggagaatg caggagctga ggaccaggat ggggagggag aaggctccaa gacagccctg
     2881 cagcctctga aacactctga cagcaaagaa gatgatggac aagaaatagc cctcgagcac
     2941 catcaccatc accatcacca tcaccattag taccaggtaa gtgtacccaa ttcgccctat
     3001 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     3061 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     3121 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     3181 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3241 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgcgactc
     3421 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4021 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5341 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5401 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     5461 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5521 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     5581 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     5641 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     5701 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5761 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5821 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5881 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5941 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6001 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6061 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6121 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6181 ttacgccagc tagagt
//
