LOCUS       sequence-470895-        4921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ATF4 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4921)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4921)
  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            "Unpublished"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/ATF4-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        125..156
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      183..192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             189..701
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             705..824
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    876..997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(887..906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     941..960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1191..1548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1191..1382
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1214..1234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1399..1534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1461..1480
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1579..2616
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2595..2603
                     /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    2640..2688
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2844..2861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3015..3603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3095..3114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3803..4594,4595..4663))
                     /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    4768..4816
                     /note="synthetic polyadenylation signal"
     misc_feature    4830..4921
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4870..4889
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgttt catcagtttc atcagtttca tcagtttcat
       61 cagtttcatc agtttcatca ctcgaggata tcaagatctg gcctcggcgg ccaagcttag
      121 acactagagg gtatataatg gaagctcgac ttccagcttg gcaatccggt actgttggta
      181 aagccaccat ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct
      241 acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg
      301 tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca
      361 tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa
      421 tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca
      481 cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag
      541 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
      601 aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca
      661 acggagtgac cggctggcgg ctgtgcgaac gcattctggc gaattctcac ggctttccgc
      721 ctgaggttga agagcaagcc gccggtacat tgcctatgtc ctgcgcacaa gaaagcggta
      781 tggaccggca cccagccgct tgtgcttcag ctcgcatcaa cgtctaaggc cgcgactcta
      841 gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
      901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1021 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1081 atgtggtaaa atcgataagg atccgtttgc gtattgggcg ctcttccgct gatctgcgca
     1141 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt tagctacctt ctgaggcgga
     1201 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     1501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct
     1561 tctgacacta gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt
     1621 ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc
     1681 cgagccttca gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct
     1741 gatggcttct acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc
     1801 cccgaagtgt tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc
     1861 gcccaaggcg ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct
     1921 gtcgccgaag ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc
     1981 ccattcgggc cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt
     2041 gctgatcccc atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta
     2101 gctcaagccc tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc
     2161 gtccatgccg acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta
     2221 atcgactggt ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc
     2281 tggcggccct ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc
     2341 gagctggccg gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc
     2401 taccagagcc tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat
     2461 gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca
     2521 gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt
     2581 acacgaccgc gcgctaagga ggtaggtcga gtttaaactc tagaaccggt catggccgca
     2641 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgtt cgaactagat
     2701 gctgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     2761 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     2821 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc
     3781 aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga
     3841 tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc
     3901 gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg
     3961 cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg
     4021 ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt
     4081 cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct
     4141 cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat
     4201 cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta
     4261 agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca
     4321 tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt
     4381 agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac
     4441 atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa
     4501 ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt
     4561 cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg
     4621 caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat
     4681 attattgaag catttatcag ggttactagt acgtctctca aggataagta agtaatatta
     4741 aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt
     4801 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     4861 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     4921 t
//
