LOCUS       sequence-471822-        6058 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 6058)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6058)
  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            "Unpublished"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: pGL4 [luc2P/ATF4-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6058
                     /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..1838
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1842..1961
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2013..2134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2024..2043)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2078..2097
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2328..2685
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2328..2519
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2351..2371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2536..2671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2598..2617
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2716..3753
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3732..3740
                     /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    3777..3825
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3981..3998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4152..4740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4232..4251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4940..5731,5732..5800))
                     /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    5905..5953
                     /note="synthetic polyadenylation signal"
     misc_feature    5967..6058
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6007..6026
                     /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 ggaagatgcc aaaaacatta agaagggccc agcgccattc tacccactcg
      241 aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg
      301 gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg
      361 agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga
      421 tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca
      481 tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca
      541 tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca
      601 acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact
      661 accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg
      721 agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca
      781 gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc
      841 gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc
      901 tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct
      961 gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc
     1021 aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga
     1081 gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc
     1141 cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc
     1201 agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa ggggacgaca
     1261 agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg gacttggaca
     1321 ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca
     1381 tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag gacggctggc
     1441 tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc gtggaccggc
     1501 tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg gagagcatcc
     1561 tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg
     1621 gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga
     1681 tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt
     1741 tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag atccgcgaga
     1801 ttctcattaa ggccaagaag ggcggcaaga tcgccgtgaa ttctcacggc ttccctcccg
     1861 aggtggagga gcaggccgcc ggcaccctgc ccatgagctg cgcccaggag agcggcatgg
     1921 atagacaccc tgctgcttgc gccagcgcca ggatcaacgt ctaaggccgc gactctagag
     1981 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2041 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2101 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2161 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2221 tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat ctgcgcagca
     2281 ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg aggcggaaag
     2341 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2401 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2641 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     2701 gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc
     2761 atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga
     2821 gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat
     2881 ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc
     2941 gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc
     3001 caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc
     3061 gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca
     3121 ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct
     3181 gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct
     3241 caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc
     3301 catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc
     3361 gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg
     3421 cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag
     3481 ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac
     3541 cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc
     3601 atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc
     3661 gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca
     3721 cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag aaccggtcat ggccgcaata
     3781 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttcga actagatgct
     3841 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     3901 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     3961 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4321 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4561 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa
     4921 tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct
     4981 gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg
     5041 agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc
     5101 ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta
     5161 ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc
     5221 cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt
     5281 cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac
     5341 ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt
     5401 tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc
     5461 catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt
     5521 gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata
     5581 gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga
     5641 tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag
     5701 catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa
     5761 agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt
     5821 attgaagcat ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg
     5881 tacgggaggt attggacagg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     5941 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     6001 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctct
//
