LOCUS       sequence-464231-        5464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses CD80 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  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 5464)
  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-CD80
FEATURES             Location/Qualifiers
     source          1..5464
                     /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"
     regulatory      1330..1339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2206..2235
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2264..2282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2556..2677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2593..2612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2647..2666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2705..2722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2876..3464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2956..2975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3635..4426,4427..4495))
                     /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     4258..4277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4496..4600)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4668..4686
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4724..4746)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4846..4865
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4932..5387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5069..5090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5281..5300
                     /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 cgcggatccg ccaccatggg ccacacacgg aggcagggaa catcaccatc caagtgtcca
     1381 tacctcaatt tctttcagct cttggtgctg gctggtcttt ctcacttctg ttcaggtgtt
     1441 atccacgtga ccaaggaagt gaaagaagtg gcaacgctgt cctgtggtca caatgtttct
     1501 gttgaagagc tggcacaaac tcgcatctac tggcaaaagg agaagaaaat ggtgctgact
     1561 atgatgtctg gggacatgaa tatatggccc gagtacaaga accggaccat ctttgatatc
     1621 actaataacc tctccattgt gatcctggct ctgcgcccat ctgacgaggg cacatacgag
     1681 tgtgttgttc tgaagtatga aaaagacgct ttcaagcggg aacacctggc tgaagtgacg
     1741 ttatcagtca aagctgactt ccctacacct agtatatctg actttgaaat tccaacttct
     1801 aatattagaa ggataatttg ctcaacctct ggaggttttc cagagcctca cctctcctgg
     1861 ttggaaaatg gagaagaatt aaatgccatc aacacaacag tttcccaaga tcctgaaact
     1921 gagctctatg ctgttagcag caaactggat ttcaatatga caaccaacca cagcttcatg
     1981 tgtctcatca agtatggaca tttaagagtg aatcagacct tcaactggaa tacaaccaag
     2041 caagagcatt ttcctgataa cctgctccca tcctgggcca ttaccttaat ctcagtaaat
     2101 ggaatttttg tgatatgctg cctgacctac tgctttgccc caagatgcag agagagaagg
     2161 aggaatgaga gattgagaag ggaaagtgta cgccctgtac tcgagcacca tcaccatcac
     2221 catcaccatc accattagta ccaggtaagt gtacccaatt cgccctatag tgagtcgtat
     2281 tacaattcac tcgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggagatcc
     2341 aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc
     2401 acagtcccaa ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca
     2461 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2521 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2581 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2641 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgcgactcta gactcttccg
     2701 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2761 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2821 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2881 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2941 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3001 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3061 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3121 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3181 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3241 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3301 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3361 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3421 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3481 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3601 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3661 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3721 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3781 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3841 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3901 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3961 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4021 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4081 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4141 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4201 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4261 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4321 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4381 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4441 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4501 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4561 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4621 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4681 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4741 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4801 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4861 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4921 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     5401 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagcta
     5461 gagt
//
