LOCUS       sequence-240495-        3677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for constitutive expression of NLuc in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3677)
  AUTHORS   Sarrion-Perdigones A, Chang L, Gonzalez Y, Gallego-Flores T, Young 
            DW, Venken KJT
  TITLE     Examining multiple cellular pathways at once using multiplex 
            hextuple luciferase assaying.
  JOURNAL   Nat Commun. 2019 Dec 13;10(1):5710. doi: 10.1038/s41467-019-13651-y.
  PUBMED    31836712
REFERENCE   2  (bases 1 to 3677)
  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: "Nat 
            Commun."; date: "2019-12-13"; volume: "10(1):5710. doi"; pages: " 
            10.1038/s41467-019-13651-y"
COMMENT     Sequence Label: hCMV-IE1:NLuc
FEATURES             Location/Qualifiers
     source          1..3677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        231..434
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     385..405
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    984..1208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1328..1916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1408..1427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2031..2133
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2134..2949
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2134..2949
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2207..2226)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3063..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3150..3169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3360..3381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        join(3604..3677,1..230)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
       61 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      121 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      181 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      241 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      301 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      361 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      421 atataagcag agctggttta gtgaaccgtc agatcaagct caaatggtgt ttaccttgga
      481 ggacttcgtg ggtgactgga ggcaaactgc aggatacaat ttggaccagg tcctcgaaca
      541 gggaggagtc agcagccttt tccaaaactt gggcgtgtcc gttaccccta ttcagagaat
      601 tgtcctttcc ggagagaacg gattgaaaat agatatccac gtcataatcc cgtatgaagg
      661 actgtctggg gaccaaatgg gtcagattga aaaaatattc aaagtagttt atcccgttga
      721 tgatcatcac ttcaaagtta tccttcatta tgggactttg gtaatcgatg gagttactcc
      781 taatatgata gattactttg gccgacctta cgaggggatc gctgtattcg acggcaagaa
      841 aatcaccgtc actggaaccc tctggaacgg caacaagatt attgacgagc gattgatcaa
      901 tccagacgga tctttgcttt tccgcgtaac gataaacggc gtaacagggt ggaggctctg
      961 cgaacgaata ctcgcctgag cttctgtgcc ttctagttgc cagccatctg ttgtttgccc
     1021 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1081 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1141 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1201 ctctatggcg cttgagacga agcttgacac ggttatccac agaatcaggg gataacgcag
     1261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac
     2041 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     2101 ttatcgagat tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct
     2161 tgctcaaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2221 cgcgataatg tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg
     2281 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2341 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2401 actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta
     2461 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2521 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgactc
     2581 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2641 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2701 ccggattcag tagtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2761 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2821 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2881 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2941 tttttctgat tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     3001 aataagtgat aataagcgga tgaatggcag aaattcgatg ataagctgtc aaacatgatg
     3061 ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3301 gccagcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat tttaacaaaa tattaacgct tacaatttta acaagcttcg tctcagtcag
     3541 gagtagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     3601 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     3661 attgacgtca ataatga
//
