LOCUS       sequence-471833-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CEBP response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4952)
  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            "Unpublished"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/CEBP-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        156..187
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      214..223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             220..732
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             736..855
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    907..1028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(918..937)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     972..991
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1222..1579
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1222..1413
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1245..1265)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1430..1565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1492..1511
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1610..2647
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2626..2634
                     /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    2671..2719
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2875..2892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3046..3634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3126..3145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3834..4625,4626..4694))
                     /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    4799..4847
                     /note="synthetic polyadenylation signal"
     misc_feature    4861..4952
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4901..4920
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcacag attgcgcaat ctaacagatt gcgcaatcta
       61 acagattgcg caatctaaca gattgcgcaa tctaacagat tgcgcaatct actcgaggat
      121 atcaagatct ggcctcggcg gccaagctta gacactagag ggtatataat ggaagctcga
      181 cttccagctt ggcaatccgg tactgttggt aaagccacca tggtcttcac actcgaagat
      241 ttcgttgggg actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga
      301 ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc
      361 ctgagcggtg aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg
      421 agcggcgacc aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat
      481 catcacttta aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac
      541 atgatcgact atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc
      601 actgtaacag ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc
      661 gacggctccc tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa
      721 cgcattctgg cgaattctca cggctttccg cctgaggttg aagagcaagc cgccggtaca
      781 ttgcctatgt cctgcgcaca agaaagcggt atggaccggc acccagccgc ttgtgcttca
      841 gctcgcatca acgtctaagg ccgcgactct agagtcgggg cggccggccg cttcgagcag
      901 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      961 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1021 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1081 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtttg
     1141 cgtattgggc gctcttccgc tgatctgcgc agcaccatgg cctgaaataa cctctgaaag
     1201 aggaacttgg ttagctacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     1261 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1321 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     1381 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     1441 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     1501 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     1561 aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc
     1621 cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga
     1681 cctgatgcag ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg
     1741 cggctatgta ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta
     1801 ccgccacttc gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag
     1861 cgagagcctg acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc
     1921 cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc
     1981 cgccgacctc agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta
     2041 cacaacctgg cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac
     2101 cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg
     2161 ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt
     2221 cctgaccgac aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga
     2281 cagtcagtac gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca
     2341 gcagactcgc tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc
     2401 ctacatgctg cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga
     2461 cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt
     2521 cggtcgcaca caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt
     2581 gctggccgac agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg
     2641 agtttaaact ctagaaccgg tcatggccgc aataaaatat ctttattttc attacatctg
     2701 tgtgttggtt ttttgtgtgt tcgaactaga tgctgtcgac cgatgccctt gagagccttc
     2761 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2821 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     2881 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3481 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3781 atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag
     3841 tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc
     3901 tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca
     3961 gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca
     4021 gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg
     4081 agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg
     4141 gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct
     4201 ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc
     4261 tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta
     4321 atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc
     4381 ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc
     4441 ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc
     4501 gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg
     4561 atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg
     4621 gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa
     4681 tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag
     4741 tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa
     4801 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     4861 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     4921 cagtgcaagt gcaggtgcca gaacatttct ct
//
