LOCUS       sequence-473374-        4942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MRE in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4942)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4942)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pNL [NlucP/MRE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        146..177
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      204..213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             210..722
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             726..845
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    897..1018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(908..927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     962..981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1212..1569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1212..1403
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1235..1255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1420..1555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1482..1501
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1600..2637
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2616..2624
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2661..2709
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2865..2882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3036..3624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3116..3135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4789..4837
                     /note="synthetic polyadenylation signal"
     misc_feature    4851..4942
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4891..4910
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgtct gcacacggcc cgtctgcaca cggcccgtct
       61 gcacacggcc cgtctgcaca cggcccgtct gcacacggcc cctcgaggat atcaagatct
      121 ggcctcggcg gccaagctta gacactagag ggtatataat ggaagctcga cttccagctt
      181 ggcaatccgg tactgttggt aaagccacca tggtcttcac actcgaagat ttcgttgggg
      241 actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca
      301 gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg
      361 aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc
      421 aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta
      481 aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact
      541 atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag
      601 ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc
      661 tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg
      721 cgaattctca cggctttccg cctgaggttg aagagcaagc cgccggtaca ttgcctatgt
      781 cctgcgcaca agaaagcggt atggaccggc acccagccgc ttgtgcttca gctcgcatca
      841 acgtctaagg ccgcgactct agagtcgggg cggccggccg cttcgagcag acatgataag
      901 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      961 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1021 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1081 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtttg cgtattgggc
     1141 gctcttccgc tgatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     1201 ttagctacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     1261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1321 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1381 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     1441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     1501 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     1561 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     1621 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     1681 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     1741 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     1801 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     1861 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     1921 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     1981 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     2041 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     2101 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     2161 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     2221 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     2281 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     2341 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     2401 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     2461 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     2521 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     2581 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttaaact
     2641 ctagaaccgg tcatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     2701 ttttgtgtgt tcgaactaga tgctgtcgac cgatgccctt gagagccttc aacccagtca
     2761 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     2821 tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg
     2881 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3721 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3781 acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag tgcttgatca
     3841 gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc tgactccccg
     3901 tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca gcaatgatgc
     3961 cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca gcagggaggg
     4021 ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg agctgctgtc
     4081 gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg gccattgcta
     4141 ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct ggttcccagc
     4201 ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc tccttagggc
     4261 ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta atggcagcac
     4321 tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc ggcgagtact
     4381 caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta
     4441 tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc gggaatcgtt
     4501 cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg atatagccca
     4561 ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa
     4621 aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc
     4681 tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag tacgtctctc
     4741 aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa taaaatatct
     4801 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     4861 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     4921 gcaggtgcca gaacatttct ct
//
