LOCUS       sequence-464229-        5551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nluc-tagged modified CG_ in mammalian cells; lectin-captured 
            luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  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 5551)
  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-Nluc-_(PLRSKK)-CA1-19
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          670..1242
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1250..1269
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1414..1923
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             2257..2286
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2350..2368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2642..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2733..2752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2791..2808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2962..3550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3042..3061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3721..4512,4513..4581))
                     /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     4344..4363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4582..4686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4754..4772
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4810..4832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4932..4951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5018..5473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5155..5176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5367..5386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      601 ggaacggtgc attggaacgg acccctaggc ttttgcaaaa agctccggat cgatcctgag
      661 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
      721 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
      781 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
      841 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
      901 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
      961 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
     1021 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
     1081 ggaaatattc ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta
     1141 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
     1201 ccctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1261 gttattgtgc tgtctcatca ttttggcaaa gaattgtaat acgactcact aagggcggcc
     1321 gccaccatga actccttctc cacaagcgcc ttcggtccag ttgccttctc cctgggcctg
     1381 ctcctggtgt tgcctgctgc cttccctgcc ccagtcttca cactcgaaga tttcgttggg
     1441 gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc
     1501 agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt
     1561 gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac
     1621 caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt
     1681 aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac
     1741 tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca
     1801 gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc
     1861 ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg
     1921 gcggctcctg atgtgcagga ttgccccgag tgcacactgc aggagaaccc ttttttttcc
     1981 cagcccggcg cccccatcct gcagtgtatg ggctgctgtt tctccagggc ctaccctacc
     2041 cctctgagga gcaagaagac catgctggtg cagaagaacg tgacaagcga gtccacatgc
     2101 tgcgtggcca agagctacaa tagggtgacc gtgatgggcg gcttcaaggt ggagaaccac
     2161 accgcctgcc actgcagcac ctgttactac cacaagtccc tgaggagatt catcgagcag
     2221 aagatcacaa agagaaagaa ggagaagtac tggcctgagc agaagctgat cagcgaggag
     2281 gatctgtgac tcgagggggg gcccggtacc ttaattaatt aaggtaccag gtaagtgtac
     2341 ccaattcgcc ctatagtgag tcgtattaca attcactcga tcgcccttcc caacagttgc
     2401 gcagcctgaa tggcgaatgg agatccaatt tttaagtgta taatgtgtta aactactgat
     2461 tctaattgtt tgtgtatttt agattcacag tcccaaggct catttcaggc ccctcagtcc
     2521 tcacagtctg ttcatgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2581 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2641 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2761 ttaacgcgcg actctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3361 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4681 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4741 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4801 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4861 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4921 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4981 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     5041 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5101 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5161 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5221 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5281 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5341 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5401 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5461 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5521 gggcctcttc gctattacgc cagctagagt c
//
