LOCUS       sequence-464284-        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nanoluc luciferase expression driven CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  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 5169)
  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-Nanoluc (Nluc)
FEATURES             Location/Qualifiers
     source          1..5169
                     /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"
     primer_bind     complement(1361..1377)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1378)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1398..1907
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             1908..1937
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1968..1986)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2260..2381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2297..2316)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2351..2370
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2409..2426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2660..2679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3339..4130,4131..4199))
                     /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     3962..3981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4372..4390
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4428..4450)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4550..4569
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4636..5091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4773..4794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4985..5004
                     /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 gcggatcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt
     1381 ctggtggatc aggtggagtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     1441 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     1501 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     1561 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     1621 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     1681 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     1741 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     1801 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     1861 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcggag cagaagctga
     1921 tcagcgagga ggatctgtag ggtaccaggt aagtgtaccc aattcgccct atagtgagtc
     1981 gtattacaat tcactcgatc gcccttccca acagttgcgc agcctgaatg gcgaatggag
     2041 atccaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag
     2101 attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata
     2161 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2221 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2341 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgcgac tctagactct
     2401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4321 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4381 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4441 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4501 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4561 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4621 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4681 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4741 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4801 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4861 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     5101 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5161 gctagagtc
//
