LOCUS       sequence-272168-        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"
     primer_bind     11..32
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        255..558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        559..762
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     713..733
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2428..2652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2772..3360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2852..2871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3475..3577
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3578..4393
                     /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             3578..4393
                     /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(3651..3670)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4507..4793,1..169)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4594..4613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
       61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttt aacaagcttc
      181 gtctcagtca ggagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      241 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      301 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      361 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      421 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      481 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      541 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      601 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      661 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      721 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      781 atcaagctca aatgcctaac gaaatcatac tccacggggc aaagccccga gatcccctcg
      841 acctcgggac ggcgggtatc cagttgtatc gcgctctgac taacttctcc tttttgcggg
      901 aggccttgat tgatgcgcac accgaagaag ttgttagtta tgcggatata ttggaaaaca
      961 gctgccgact tgccaaatgc tatgagaatt acggcctgcg ccaaaactca gtgataagtg
     1021 tttgctccga aaactcaaca atttttttct accccgtgat tgctgccctc tatatgggtg
     1081 taattactgc cactgtgaat gactcttata ccgaacgcga attgcttgaa acgctcaaca
     1141 tttcaaagcc ggagttggta ttttgcagta aaaaagcaat caagaacatg atggctctca
     1201 aaagaaatgt aaactttata aagaaagtcg tgctcctcga ttccaaggag gatatgggcg
     1261 aggcccaatg tctgagcaac tttatggctc ggtattccga accaaatttg gacgttcgga
     1321 attttaaacc ccgcgacttt gatgcaaagg aacaagttgc tctcattatg tcctcttccg
     1381 ggaccaccgg acttccaaaa ggagtggtac tcacgcacag aaacctctca gtaagatttg
     1441 ttcactgcaa ggatccactt ttcgggacaa ggacaatacc tagtacttcc attctgagta
     1501 tagtgccatt tcatcatgca ttcggaatgt tcacaacatt gtcttacttc atcgtgggct
     1561 tgagggtagt actccttaag cgcttcgaag agaaattttt cctgtctact atcgagaaat
     1621 accgcattcc gaccatcgtg cttgcgccac cagttatggt attcctcgcc aaatccccgt
     1681 tggtagacca gtatgacttg tcctctatac gcgaggttgc caccggcggt gccccagtcg
     1741 ggaccgaagt agccgttgcc gtagcgaaac ggctgaaaat cggcggcatt ctgcaaggtt
     1801 atggactcac ggaaacgtgt tgtgcggtgc tcatcacgcc gcacgacgac gtcaagaccg
     1861 gctcaacagg ccgggttgcg ccatacgtac aagcaaagat agtagatctt accacgggaa
     1921 aatctctggg gcctaacaag agaggtgaac tgtgcttcaa aagtgaaatt attatgaagg
     1981 gctactttaa taataaacaa gccaccgaag aagccattga taaggaaggg tggctccact
     2041 ctggggatgt cgggtactac gacgatgatg gccacttttt tgtcgttgat cggcttaagg
     2101 agctgatcaa gtacaagggt tatcaagtcg ccccagccga gcttgagtgg ctcctcctcc
     2161 aacatcccag cattaaggac gcaggagtaa ctggggtgcc cgatgaggca gccggcgagt
     2221 tgccgggggc atgtattgtg ttgcaagaag gaaaatcatt gaccgaacaa gagatcatag
     2281 attacattgc ggaaagagtt agccccacaa aacgcatccg agggggagta gttttcgtag
     2341 acgatatacc taaaggcgcg acaggcaagt tggttcgatc cgaattgcgc aaacttctcg
     2401 cgcagaagaa atccaagttg tgagcttctg tgccttctag ttgccagcca tctgttgttt
     2461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2641 tgggctctat ggcgcttgag acgaagcttg acacggttat ccacagaatc aggggataac
     2701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3181 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gtgatgatcg
     3481 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3541 tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac
     3601 gtcttgctca aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3661 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctac cgattgtatg ggaagcccga
     3721 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3781 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3841 ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca
     3901 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3961 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4021 actcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4081 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4141 ctcaccggat tcagtagtca ctcatggtga tttctcactt gataacctta tttttgacga
     4201 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4261 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4321 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4381 tgagtttttc tgattttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     4441 cctgaataag tgataataag cggatgaatg gcagaaattc gatgataagc tgtcaaacat
     4501 gatgttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4561 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4621 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4681 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4741 gtgggccagc gccctgatag acggtttttc gccctttgac gttggagtcc acg
//
