LOCUS       sequence-230608-        4097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for constitutive expression of GrRenilla in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4097)
  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 4097)
  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:GrRenilla
FEATURES             Location/Qualifiers
     source          1..4097
                     /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    1563..1787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1907..2495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1987..2006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2610..2712
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2713..3528
                     /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             2713..3528
                     /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(2786..2805)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3642..4070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3729..3748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3939..3960
                     /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 aaatggcttc caaagtctac gacccagaac aaaggaagag
      661 gatgattact ggaccgcagt ggtgggcaag gtgcaagcag atgaatgtac tcgatagttt
      721 tatcaattat tacgactccg aaaaacacgc ggaaaatgct gtcatattcc tgcacgggaa
      781 cgccactagc agctacttgt ggcggcacgt agtcccccac atagagcctg ttgcccgatg
      841 tattattccc gatctcattg gaatgggcaa atcaggcaaa tctgggaacg gttcttaccg
      901 cctgctcgat cactataaat atttgaccgc ttggtttgag ctgcttaatc tccctaaaaa
      961 aataatattc gttggacacg actggggctc agctctggcg ttccactacg catacgagca
     1021 tcaagaccga ataaaagcaa tagtacatat ggagtcagtt gttgacgtca tagagtcatg
     1081 gatgggttgg ccagatattg aagaggaatt ggccctcatc aagagtgagg agggggaaaa
     1141 aatggtcctg gaaaataatt ttttcgtgga aaccttgctc ccgagtaaaa taatgcgaaa
     1201 acttgaaccc gaagagtttg ctgcttattt ggagcctttc aaggaaaagg gggaagtaag
     1261 acgaccaacc ctttcttggc cgagggagat acctcttgtt aagggtggaa agcccgacgt
     1321 agtccagata gtccgaaatt acaacgcgta cctgagagct agtgatgacc tccctaagtt
     1381 gtttatcgag tcagaccctg ggttcttcag caacgcaatt gtagaaggag cgaaaaaatt
     1441 tccaaataca gaatttgtga aggttaaggg gctccacttt ttgcaagagg acgcccccga
     1501 tgagatggga aaatacatta aatctttcgt agagcgcgtc cttaagaatg aacagtgagc
     1561 ttctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1621 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1681 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1741 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggcgc ttgagacgaa
     1801 gcttgacacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2581 aaaggatctt cacctagatc cttttgtgat gatcggcacg taagaggttc caactttcac
     2641 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     2701 aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat
     2761 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2821 ggtgcgacaa tctaccgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2881 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2941 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3001 ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca
     3061 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3121 tgtaattgtc cttttaacag cgatcgcgta tttcgactcg ctcaggcgca atcacgaatg
     3181 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3241 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     3301 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3361 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3421 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3481 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctgatt ttttttaagg
     3541 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat
     3601 gaatggcaga aattcgatga taagctgtca aacatgatgt tacgcgccct gtagcggcgc
     3661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3841 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt
     3901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3961 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4021 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4081 attaacgctt acaattt
//
