LOCUS       sequence-464289-        5506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses ICOSLG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5506)
  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 5506)
  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-ICOSLG
FEATURES             Location/Qualifiers
     source          1..5506
                     /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             2248..2277
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2306..2324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2598..2719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2635..2654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2689..2708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2747..2764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2918..3506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2998..3017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3677..4468,4469..4537))
                     /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     4300..4319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4538..4642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4710..4728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4766..4788)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4888..4907
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4974..5429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5111..5132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5323..5342
                     /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 ccaccatgcg gctgggcagt cctggactgc tcttcctgct cttcagcagc
     1381 cttcgagctg atactcagga gaaggaagtc agagcgatgg taggcagcga cgtggagctc
     1441 agctgcgctt gccctgaagg aagccgtttt gatttaaatg atgtttacgt atattggcaa
     1501 accagtgagt cgaaaaccgt ggtgacctac cacatcccac agaacagctc cttggaaaac
     1561 gtggacagcc gctaccggaa ccgagccctg atgtcaccgg ccggcatgct gcggggcgac
     1621 ttctccctgc gcttgttcaa cgtcaccccc caggacgagc agaagtttca ctgcctggtg
     1681 ttgagccaat ccctgggatt ccaggaggtt ttgagcgttg aggttacact gcatgtggca
     1741 gcaaacttca gcgtgcccgt cgttagcgcc ccccacagcc cctcccagga tgagctcacc
     1801 ttcacgtgta catccataaa cggctacccc aggcccaacg tgtactggat caataagacg
     1861 gacaacagcc tgctggacca ggctctgcag aatgacaccg tcttcttgaa catgcggggc
     1921 ttgtatgacg tggtcagcgt gctgaggatc gcacggaccc ccagcgtgaa cattggctgc
     1981 tgcatagaga acgtgcttct gcagcagaac ctgactgtcg gcagccagac aggaaatgac
     2041 atcggagaga gagacaagat cacagagaat ccagtcagta ccggcgagaa aaacgcggcc
     2101 acgtggagca tcctggctgt cctgtgcctg cttgtggtcg tggcggtggc cataggctgg
     2161 gtgtgcaggg accgatgcct ccaacacagc tatgcaggtg cctgggctgt gagtccggag
     2221 acagagctca ctggccacgt tctcgagcac catcaccatc accatcacca tcaccattag
     2281 taccaggtaa gtgtacccaa ttcgccctat agtgagtcgt attacaattc actcgatcgc
     2341 ccttcccaac agttgcgcag cctgaatggc gaatggagat ccaattttta agtgtataat
     2401 gtgttaaact actgattcta attgtttgtg tattttagat tcacagtccc aaggctcatt
     2461 tcaggcccct cagtcctcac agtctgttca tgatcataat cagccatacc acatttgtag
     2521 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2581 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2641 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2701 aactcatcaa tgtatcttaa cgcgcgactc tagactcttc cgcttcctcg ctcactgact
     2761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4681 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     4741 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4801 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4861 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4921 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     4981 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5041 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5101 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5161 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5221 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5281 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5341 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5401 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     5461 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tagagt
//
