LOCUS       sequence-261436-        4793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for constitutive expression of RoLuc in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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 4793)
  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:RoLuc:bgH
FEATURES             Location/Qualifiers
     source          1..4793
                     /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    2259..2483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2603..3191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2683..2702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3306..3408
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3409..4224
                     /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             3409..4224
                     /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(3482..3501)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4338..4766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4425..4444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4635..4656
                     /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 aaatgcctaa cgaaatcata ctccacgggg caaagccccg
      661 agatcccctc gacctcggga cggcgggtat ccagttgtat cgcgctctga ctaacttctc
      721 ctttttgcgg gaggccttga ttgatgcgca caccgaagaa gttgttagtt atgcggatat
      781 attggaaaac agctgccgac ttgccaaatg ctatgagaat tacggcctgc gccaaaactc
      841 agtgataagt gtttgctccg aaaactcaac aatttttttc taccccgtga ttgctgccct
      901 ctatatgggt gtaattactg ccactgtgaa tgactcttat accgaacgcg aattgcttga
      961 aacgctcaac atttcaaagc cggagttggt attttgcagt aaaaaagcaa tcaagaacat
     1021 gatggctctc aaaagaaatg taaactttat aaagaaagtc gtgctcctcg attccaagga
     1081 ggatatgggc gaggcccaat gtctgagcaa ctttatggct cggtattccg aaccaaattt
     1141 ggacgttcgg aattttaaac cccgcgactt tgatgcaaag gaacaagttg ctctcattat
     1201 gtcctcttcc gggaccaccg gacttccaaa aggagtggta ctcacgcaca gaaacctctc
     1261 agtaagattt gttcactgca aggatccact tttcgggaca aggacaatac ctagtacttc
     1321 cattctgagt atagtgccat ttcatcatgc attcggaatg ttcacaacat tgtcttactt
     1381 catcgtgggc ttgagggtag tactccttaa gcgcttcgaa gagaaatttt tcctgtctac
     1441 tatcgagaaa taccgcattc cgaccatcgt gcttgcgcca ccagttatgg tattcctcgc
     1501 caaatccccg ttggtagacc agtatgactt gtcctctata cgcgaggttg ccaccggcgg
     1561 tgccccagtc gggaccgaag tagccgttgc cgtagcgaaa cggctgaaaa tcggcggcat
     1621 tctgcaaggt tatggactca cggaaacgtg ttgtgcggtg ctcatcacgc cgcacgacga
     1681 cgtcaagacc ggctcaacag gccgggttgc gccatacgta caagcaaaga tagtagatct
     1741 taccacggga aaatctctgg ggcctaacaa gagaggtgaa ctgtgcttca aaagtgaaat
     1801 tattatgaag ggctacttta ataataaaca agccaccgaa gaagccattg ataaggaagg
     1861 gtggctccac tctggggatg tcgggtacta cgacgatgat ggccactttt ttgtcgttga
     1921 tcggcttaag gagctgatca agtacaaggg ttatcaagtc gccccagccg agcttgagtg
     1981 gctcctcctc caacatccca gcattaagga cgcaggagta actggggtgc ccgatgaggc
     2041 agccggcgag ttgccggggg catgtattgt gttgcaagaa ggaaaatcat tgaccgaaca
     2101 agagatcata gattacattg cggaaagagt tagccccaca aaacgcatcc gagggggagt
     2161 agttttcgta gacgatatac ctaaaggcgc gacaggcaag ttggttcgat ccgaattgcg
     2221 caaacttctc gcgcagaaga aatccaagtt gtgagcttct gtgccttcta gttgccagcc
     2281 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2341 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2401 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2461 tggggatgcg gtgggctcta tggcgcttga gacgaagctt gacacggtta tccacagaat
     2521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3301 tgtgatgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     3361 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat gagccatatt
     3421 caacgggaaa cgtcttgctc aaggccgcga ttaaattcca acatggatgc tgatttatat
     3481 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta ccgattgtat
     3541 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     3601 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     3661 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa
     3721 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     3781 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     3841 cgcgtatttc gactcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     3901 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     3961 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4021 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4081 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4141 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4201 ttgatgctcg atgagttttt ctgatttttt ttaaggcagt tattggtgcc cttaaacgcc
     4261 tggttgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgatgataag
     4321 ctgtcaaaca tgatgttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4561 tggttcacgt agtgggccag cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa ttt
//
