LOCUS       sequence-472870-        4949 bp    DNA     circular SYN 14-MAY-2025
DEFINITION  Express ARE response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4949)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4949)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL [NlucP/ARE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        153..184
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      211..220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             217..729
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             733..852
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    904..1025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(915..934)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     969..988
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1219..1576
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1219..1410
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1242..1262)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1427..1562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1489..1508
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1607..2644
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2623..2631
                     /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    2668..2716
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2872..2889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3043..3631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3123..3142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3831..4622,4623..4691))
                     /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    4796..4844
                     /note="synthetic polyadenylation signal"
     misc_feature    4858..4949
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4898..4917
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctctagc ttggaaatga cattgctaat ggtgacaaag
       61 caacttttag cttggaaatg acattgctaa tggtgacaaa gcaactttct cgaggatatc
      121 aagatctggc ctcggcggcc aagcttagac actagagggt atataatgga agctcgactt
      181 ccagcttggc aatccggtac tgttggtaaa gccaccatgg tcttcacact cgaagatttc
      241 gttggggact ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt
      301 gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg
      361 agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc
      421 ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat
      481 cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg
      541 atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact
      601 gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac
      661 ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc
      721 attctggcga attctcacgg ctttccgcct gaggttgaag agcaagccgc cggtacattg
      781 cctatgtcct gcgcacaaga aagcggtatg gaccggcacc cagccgcttg tgcttcagct
      841 cgcatcaacg tctaaggccg cgactctaga gtcggggcgg ccggccgctt cgagcagaca
      901 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      961 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1021 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1081 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtttgcgt
     1141 attgggcgct cttccgctga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     1201 aacttggtta gctaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     1261 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1321 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1381 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     1441 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     1501 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgagtaggct tttttggagg
     1561 cctaggcttt tgcaaaaagc tcgattcttc tgacactagc gccaccatga agaagcccga
     1621 actcaccgct accagcgttg aaaaatttct catcgagaag ttcgacagtg tgagcgacct
     1681 gatgcagttg tcggagggcg aagagagccg agccttcagc ttcgatgtcg gcggacgcgg
     1741 ctatgtactg cgggtgaata gctgcgctga tggcttctac aaagaccgct acgtgtaccg
     1801 ccacttcgcc agcgctgcac tacccatccc cgaagtgttg gacatcggcg agttcagcga
     1861 gagcctgaca tactgcatca gtagacgcgc ccaaggcgtt actctccaag acctccccga
     1921 aacagagctg cctgctgtgt tacagcctgt cgccgaagct atggatgcta ttgccgccgc
     1981 cgacctcagt caaaccagcg gcttcggccc attcgggccc caaggcatcg gccagtacac
     2041 aacctggcgg gatttcattt gcgccattgc tgatccccat gtctaccact ggcagaccgt
     2101 gatggacgac accgtgtccg ccagcgtagc tcaagccctg gacgaactga tgctgtgggc
     2161 cgaagactgt cccgaggtgc gccacctcgt ccatgccgac ttcggcagca acaacgtcct
     2221 gaccgacaac ggccgcatca ccgccgtaat cgactggtcc gaagctatgt tcggggacag
     2281 tcagtacgag gtggccaaca tcttcttctg gcggccctgg ctggcttgca tggagcagca
     2341 gactcgctac ttcgagcgcc ggcatcccga gctggccggc agccctcgtc tgcgagccta
     2401 catgctgcgc atcggcctgg atcagctcta ccagagcctc gtggacggca acttcgacga
     2461 tgctgcctgg gctcaaggcc gctgcgatgc catcgtccgc agcggggccg gcaccgtcgg
     2521 tcgcacacaa atcgctcgcc ggagcgcagc cgtatggacc gacggctgcg tcgaggtgct
     2581 ggccgacagc ggcaaccgcc ggcccagtac acgaccgcgc gctaaggagg taggtcgagt
     2641 ttaaactcta gaaccggtca tggccgcaat aaaatatctt tattttcatt acatctgtgt
     2701 gttggttttt tgtgtgttcg aactagatgc tgtcgaccga tgcccttgag agccttcaac
     2761 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2821 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     2881 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2941 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3481 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3601 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3721 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3781 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc
     3841 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga
     3901 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca
     3961 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca
     4021 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc
     4081 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc
     4141 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt
     4201 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc
     4261 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg
     4321 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc
     4381 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg
     4441 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg
     4501 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata
     4561 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg
     4621 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt
     4681 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac
     4741 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa
     4801 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac
     4861 atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag
     4921 tgcaagtgca ggtgccagaa catttctct
//
