LOCUS       sequence-474661-        4908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HSE in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4908)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4908)
  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            "Unpublished"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/HSE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        112..143
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      170..179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             176..688
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             692..811
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    863..984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(874..893)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     928..947
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1178..1535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1178..1369
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1201..1221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1386..1521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1448..1467
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1566..2603
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2582..2590
                     /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    2627..2675
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2831..2848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3002..3590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3082..3101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /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    4755..4803
                     /note="synthetic polyadenylation signal"
     misc_feature    4817..4908
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4857..4876
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcctgg aagattctag aacgttctgg aagattctag
       61 aacgttcctc gaggatatca agatctggcc tcggcggcca agcttagaca ctagagggta
      121 tataatggaa gctcgacttc cagcttggca atccggtact gttggtaaag ccaccatggt
      181 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
      241 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
      301 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
      361 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
      421 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
      481 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
      541 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
      601 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
      661 ctggcggctg tgcgaacgca ttctggcgaa ttctcacggc tttccgcctg aggttgaaga
      721 gcaagccgcc ggtacattgc ctatgtcctg cgcacaagaa agcggtatgg accggcaccc
      781 agccgcttgt gcttcagctc gcatcaacgt ctaaggccgc gactctagag tcggggcggc
      841 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
      901 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
      961 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1021 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     1081 gataaggatc cgtttgcgta ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg
     1141 aaataacctc tgaaagagga acttggttag ctaccttctg aggcggaaag aaccagctgt
     1201 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1261 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     1321 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     1381 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     1441 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     1501 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct gacactagcg
     1561 ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc atcgagaagt
     1621 tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga gccttcagct
     1681 tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat ggcttctaca
     1741 aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc gaagtgttgg
     1801 acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc caaggcgtta
     1861 ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc gccgaagcta
     1921 tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca ttcgggcccc
     1981 aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct gatccccatg
     2041 tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct caagccctgg
     2101 acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc catgccgact
     2161 tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc gactggtccg
     2221 aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg cggccctggc
     2281 tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag ctggccggca
     2341 gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac cagagcctcg
     2401 tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca
     2461 gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg
     2521 acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg
     2581 ctaaggaggt aggtcgagtt taaactctag aaccggtcat ggccgcaata aaatatcttt
     2641 attttcatta catctgtgtg ttggtttttt gtgtgttcga actagatgct gtcgaccgat
     2701 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2761 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3421 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3721 aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa tgctaaacca
     3781 ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct gcctatttcg
     3841 ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg agggcttacc
     3901 atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc
     3961 agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc
     4021 ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc cagtgagtag
     4081 tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt cgttcggtat
     4141 ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac ccatattatg
     4201 aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt tggccgcggt
     4261 gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc catccgtaag
     4321 atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt gtatacggcg
     4381 accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata gcagtacttt
     4441 gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga tcttgccgct
     4501 attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag catcttttac
     4561 tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat
     4621 gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt attgaagcat
     4681 ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt
     4741 attggacagg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     4801 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     4861 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctct
//
