LOCUS       sequence-464230-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses VTCN1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  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 5476)
  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-VTCN1
FEATURES             Location/Qualifiers
     source          1..5476
                     /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             1408..1437
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2218..2247
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(2276..2294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2568..2689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2605..2624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2659..2678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2717..2734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2888..3476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2968..2987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3647..4438,4439..4507))
                     /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     4270..4289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4508..4612)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4680..4698
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4736..4758)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4858..4877
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4944..5399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5081..5102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5293..5312
                     /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 ccaccatggc ttccctgggg cagatcctct tctggagcat aattagcatc
     1381 atcattattc tggctggagc aattgcacac catcaccatc accatcacca tcaccatctc
     1441 atcattggct ttggtatttc agggagacac tccatcacag tcactactgt cgcctcagct
     1501 gggaacattg gggaggatgg aatcctgagc tgcacttttg aacctgacat caaactttct
     1561 gatatcgtga tacaatggct gaaggaaggt gttttaggct tggtccatga gttcaaagaa
     1621 ggcaaagatg agctgtcgga gcaggatgaa atgttcagag gccggacagc agtgtttgct
     1681 gatcaagtga tagttggcaa tgcctctttg cggctgaaaa acgtgcaact cacagatgct
     1741 ggcacctaca aatgttatat catcacttct aaaggcaagg ggaatgctaa ccttgagtat
     1801 aaaactggag ccttcagcat gccggaagtg aatgtggact ataatgccag ctcagagacc
     1861 ttgcggtgtg aggctccccg atggttcccc cagcccacag tggtctgggc atcccaagtt
     1921 gaccagggag ccaacttctc ggaagtctcc aataccagct ttgagctgaa ctctgagaat
     1981 gtgaccatga aggttgtgtc tgtgctctac aatgttacga tcaacaacac atactcctgt
     2041 atgattgaaa atgacattgc caaagcaaca ggggatatca aagtgacaga atcggagatc
     2101 aaaaggcgga gtcacctaca gctgctaaac tcaaaggctt ctctgtgtgt ctcttctttc
     2161 tttgccatca gctgggcact tctgcctctc agcccttacc tgatgctaaa actcgagcac
     2221 catcaccatc accatcacca tcaccattag taccaggtaa gtgtacccaa ttcgccctat
     2281 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     2341 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     2401 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     2461 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2521 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2581 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2641 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgcgactc
     2701 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2761 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2821 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4621 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4681 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4741 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4801 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4861 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4921 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     4981 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5041 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5101 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5161 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5221 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5281 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5341 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5401 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5461 ttacgccagc tagagt
//
