LOCUS       sequence-230547-        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"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1143..1367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1487..2075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1567..1586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2190..2292
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2293..3108
                     /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             2293..3108
                     /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(2366..2385)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3222..3650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3309..3328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3519..3540
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacaagctt cgtctcagtc aggagtagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatcaagctc aaatggtgtt taccttggag gacttcgtgg gtgactggag
      661 gcaaactgca ggatacaatt tggaccaggt cctcgaacag ggaggagtca gcagcctttt
      721 ccaaaacttg ggcgtgtccg ttacccctat tcagagaatt gtcctttccg gagagaacgg
      781 attgaaaata gatatccacg tcataatccc gtatgaagga ctgtctgggg accaaatggg
      841 tcagattgaa aaaatattca aagtagttta tcccgttgat gatcatcact tcaaagttat
      901 ccttcattat gggactttgg taatcgatgg agttactcct aatatgatag attactttgg
      961 ccgaccttac gaggggatcg ctgtattcga cggcaagaaa atcaccgtca ctggaaccct
     1021 ctggaacggc aacaagatta ttgacgagcg attgatcaat ccagacggat ctttgctttt
     1081 ccgcgtaacg ataaacggcg taacagggtg gaggctctgc gaacgaatac tcgcctgagc
     1141 ttctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1201 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1261 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1321 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggcgc ttgagacgaa
     1381 gcttgacacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1441 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1501 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1561 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1621 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1681 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1741 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1801 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1861 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1921 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1981 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2041 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2101 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2161 aaaggatctt cacctagatc cttttgtgat gatcggcacg taagaggttc caactttcac
     2221 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     2281 aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat
     2341 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2401 ggtgcgacaa tctaccgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2461 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2521 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2581 ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca
     2641 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2701 tgtaattgtc cttttaacag cgatcgcgta tttcgactcg ctcaggcgca atcacgaatg
     2761 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2821 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2881 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2941 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3001 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3061 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctgatt ttttttaagg
     3121 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat
     3181 gaatggcaga aattcgatga taagctgtca aacatgatgt tacgcgccct gtagcggcgc
     3241 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3301 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3361 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3421 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt
     3481 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3541 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3601 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3661 attaacgctt acaattt
//
