LOCUS       sequence-470903-        4943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HRE response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4943)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4943)
  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            "Unpublished"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/HRE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        147..178
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      205..214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             211..723
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             727..846
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    898..1019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(909..928)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     963..982
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1213..1570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1213..1404
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1236..1256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1421..1556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1483..1502
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1601..2638
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2617..2625
                     /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    2662..2710
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2866..2883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3037..3625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3117..3136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3825..4616,4617..4685))
                     /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    4790..4838
                     /note="synthetic polyadenylation signal"
     misc_feature    4852..4943
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4892..4911
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgtga ctacgtgctg cctaggtgac tacgtgctgc
       61 ctaggtgact acgtgctgcc taggtgacta cgtgctgcct agctcgagga tatcaagatc
      121 tggcctcggc ggccaagctt agacactaga gggtatataa tggaagctcg acttccagct
      181 tggcaatccg gtactgttgg taaagccacc atggtcttca cactcgaaga tttcgttggg
      241 gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc
      301 agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt
      361 gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac
      421 caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt
      481 aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac
      541 tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca
      601 gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc
      661 ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg
      721 gcgaattctc acggctttcc gcctgaggtt gaagagcaag ccgccggtac attgcctatg
      781 tcctgcgcac aagaaagcgg tatggaccgg cacccagccg cttgtgcttc agctcgcatc
      841 aacgtctaag gccgcgactc tagagtcggg gcggccggcc gcttcgagca gacatgataa
      901 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      961 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1021 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1081 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgttt gcgtattggg
     1141 cgctcttccg ctgatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     1201 gttagctacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     1261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1321 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1381 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     1441 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1501 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     1561 cttttgcaaa aagctcgatt cttctgacac tagcgccacc atgaagaagc ccgaactcac
     1621 cgctaccagc gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg acctgatgca
     1681 gttgtcggag ggcgaagaga gccgagcctt cagcttcgat gtcggcggac gcggctatgt
     1741 actgcgggtg aatagctgcg ctgatggctt ctacaaagac cgctacgtgt accgccactt
     1801 cgccagcgct gcactaccca tccccgaagt gttggacatc ggcgagttca gcgagagcct
     1861 gacatactgc atcagtagac gcgcccaagg cgttactctc caagacctcc ccgaaacaga
     1921 gctgcctgct gtgttacagc ctgtcgccga agctatggat gctattgccg ccgccgacct
     1981 cagtcaaacc agcggcttcg gcccattcgg gccccaaggc atcggccagt acacaacctg
     2041 gcgggatttc atttgcgcca ttgctgatcc ccatgtctac cactggcaga ccgtgatgga
     2101 cgacaccgtg tccgccagcg tagctcaagc cctggacgaa ctgatgctgt gggccgaaga
     2161 ctgtcccgag gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg tcctgaccga
     2221 caacggccgc atcaccgccg taatcgactg gtccgaagct atgttcgggg acagtcagta
     2281 cgaggtggcc aacatcttct tctggcggcc ctggctggct tgcatggagc agcagactcg
     2341 ctacttcgag cgccggcatc ccgagctggc cggcagccct cgtctgcgag cctacatgct
     2401 gcgcatcggc ctggatcagc tctaccagag cctcgtggac ggcaacttcg acgatgctgc
     2461 ctgggctcaa ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac
     2521 acaaatcgct cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg tgctggccga
     2581 cagcggcaac cgccggccca gtacacgacc gcgcgctaag gaggtaggtc gagtttaaac
     2641 tctagaaccg gtcatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     2701 tttttgtgtg ttcgaactag atgctgtcga ccgatgccct tgagagcctt caacccagtc
     2761 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     2821 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
     2881 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2941 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3001 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3061 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3121 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3181 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3241 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3301 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3361 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3421 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3481 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3541 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3601 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3661 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3721 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3781 aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca gtgcttgatc
     3841 agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc ctgactcccc
     3901 gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc agcaatgatg
     3961 ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg
     4021 gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat gagctgctgt
     4081 cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt ggccattgct
     4141 actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc tggttcccag
     4201 cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag ctccttaggg
     4261 cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt aatggcagca
     4321 ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac cggcgagtac
     4381 tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg cccggcgtct
     4441 atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat cgggaatcgt
     4501 tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc gatatagccc
     4561 actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc ggggtgtgca
     4621 aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg
     4681 ctcatactcg tcctttttca atattattga agcatttatc agggttacta gtacgtctct
     4741 caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca ataaaatatc
     4801 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     4861 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     4921 tgcaggtgcc agaacatttc tct
//
