LOCUS       sequence-471831-        4974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ATF6 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4974)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4974)
  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            "Unpublished"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/ATF6-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        178..209
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      236..245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             242..754
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             758..877
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    929..1050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(940..959)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     994..1013
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1244..1601
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1244..1435
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1267..1287)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1452..1587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1514..1533
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1632..2669
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2648..2656
                     /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    2693..2741
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2897..2914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3068..3656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3148..3167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3856..4647,4648..4716))
                     /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    4821..4869
                     /note="synthetic polyadenylation signal"
     misc_feature    4883..4974
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4923..4942
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcatcg agacaggtgc tgacgtggca ttcatcgaga
       61 caggtgctga cgtggcattc atcgagacag gtgctgacgt ggcattcatc gagacaggtg
      121 ctgacgtggc attcatcgag acaggtgctg acgtggcatt cctcgaagct tagacactag
      181 agggtatata atggaagctc gacttccagc ttggcaatcc ggtactgttg gtaaagccac
      241 catggtcttc acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct
      301 ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt
      361 aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt
      421 catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa
      481 ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt
      541 aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc
      601 cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat
      661 cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt
      721 gaccggctgg cggctgtgcg aacgcattct ggcgaattct cacggctttc cgcctgaggt
      781 tgaagagcaa gccgccggta cattgcctat gtcctgcgca caagaaagcg gtatggaccg
      841 gcacccagcc gcttgtgctt cagctcgcat caacgtctaa ggccgcgact ctagagtcgg
      901 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
      961 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1021 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1081 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1141 aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc gcagcaccat
     1201 ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc ggaaagaacc
     1261 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1321 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     1381 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     1441 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1501 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     1561 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca
     1621 ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg
     1681 agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct
     1741 tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct
     1801 tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag
     1861 tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag
     1921 gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg
     1981 aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg
     2041 ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc
     2101 cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag
     2161 ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg
     2221 ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact
     2281 ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc
     2341 cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg
     2401 ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga
     2461 gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg
     2521 tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat
     2581 ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac
     2641 cgcgcgctaa ggaggtaggt cgagtttaaa ctctagaacc ggtcatggcc gcaataaaat
     2701 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gttcgaacta gatgctgtcg
     2761 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2821 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2881 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2941 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3001 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3481 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
     3841 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
     3901 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
     3961 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
     4021 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
     4081 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
     4141 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
     4201 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
     4261 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
     4321 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
     4381 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
     4441 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
     4501 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
     4561 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
     4621 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
     4681 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
     4741 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
     4801 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4861 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     4921 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
