LOCUS       sequence-471832-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LCN2 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6145)
  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            "Unpublished"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/LCN2/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      270..279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             276..1925
                     /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             1929..2048
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2100..2221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2111..2130)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2165..2184
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2415..2772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2415..2606
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2438..2458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2623..2758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2685..2704
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2803..3840
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3819..3827
                     /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    3864..3912
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4068..4085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4239..4827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4319..4338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5027..5818,5819..5887))
                     /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    5992..6040
                     /note="synthetic polyadenylation signal"
     misc_feature    6054..6145
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6094..6113
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca gaggtgcagc actccgggaa tgtccctcac tctccccgtc
       61 cctctgtctt gcccaatcct gaccaggtgc agaaatcttg ccaagtgttt ccgcaggagt
      121 tgctggcaat tgcctcacat tcctggcctt ggcaaagaat gaatcaaccc accctagatc
      181 ccataaatag ggccacccag gtgagcctct cactcgccac ctcctcttcc acccctgcca
      241 aagcttggca atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga
      301 agggcccagc gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag
      361 ccatgaagcg ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg
      421 tggacattac ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc
      481 gctatgggct gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct
      541 tcatgcccgt gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct
      601 acaacgagcg cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga
      661 gcaagaaagg gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga
      721 tcatcatcat ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga
      781 cttcccattt gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg
      841 acaaaaccat cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag
      901 ccctaccgca ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca
      961 accagatcat ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca
     1021 tgttcaccac gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg
     1081 aggaggagct attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc
     1141 ccacactatt tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact
     1201 tgcacgagat cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca
     1261 aacgcttcca cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca
     1321 ttctgatcac ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct
     1381 tcgaggctaa ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg
     1441 agctgtgcgt ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa
     1501 acgctctcat cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg
     1561 acgagcactt cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg
     1621 tagccccagc cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg
     1681 tcgccggcct gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac
     1741 acggtaaaac catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg
     1801 ccaagaagct gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca
     1861 agttggacgc ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg
     1921 ccgtgaattc tcacggcttc cctcccgagg tggaggagca ggccgccggc accctgccca
     1981 tgagctgcgc ccaggagagc ggcatggata gacaccctgc tgcttgcgcc agcgccagga
     2041 tcaacgtcta aggccgcgac tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     2101 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2161 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2221 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2281 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt ttgcgtattg
     2341 ggcgctcttc cgctgatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     2401 tggttagcta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2701 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2761 ggcttttgca aaaagctcga ttcttctgac actagcgcca ccatgaagaa gcccgaactc
     2821 accgctacca gcgttgaaaa atttctcatc gagaagttcg acagtgtgag cgacctgatg
     2881 cagttgtcgg agggcgaaga gagccgagcc ttcagcttcg atgtcggcgg acgcggctat
     2941 gtactgcggg tgaatagctg cgctgatggc ttctacaaag accgctacgt gtaccgccac
     3001 ttcgccagcg ctgcactacc catccccgaa gtgttggaca tcggcgagtt cagcgagagc
     3061 ctgacatact gcatcagtag acgcgcccaa ggcgttactc tccaagacct ccccgaaaca
     3121 gagctgcctg ctgtgttaca gcctgtcgcc gaagctatgg atgctattgc cgccgccgac
     3181 ctcagtcaaa ccagcggctt cggcccattc gggccccaag gcatcggcca gtacacaacc
     3241 tggcgggatt tcatttgcgc cattgctgat ccccatgtct accactggca gaccgtgatg
     3301 gacgacaccg tgtccgccag cgtagctcaa gccctggacg aactgatgct gtgggccgaa
     3361 gactgtcccg aggtgcgcca cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc
     3421 gacaacggcc gcatcaccgc cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag
     3481 tacgaggtgg ccaacatctt cttctggcgg ccctggctgg cttgcatgga gcagcagact
     3541 cgctacttcg agcgccggca tcccgagctg gccggcagcc ctcgtctgcg agcctacatg
     3601 ctgcgcatcg gcctggatca gctctaccag agcctcgtgg acggcaactt cgacgatgct
     3661 gcctgggctc aaggccgctg cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc
     3721 acacaaatcg ctcgccggag cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc
     3781 gacagcggca accgccggcc cagtacacga ccgcgcgcta aggaggtagg tcgagtttaa
     3841 actctagaac cggtcatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     3901 gttttttgtg tgttcgaact agatgctgtc gaccgatgcc cttgagagcc ttcaacccag
     3961 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     4021 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     4081 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4141 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4201 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4261 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4321 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4381 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4441 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4501 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4561 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4621 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4681 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4741 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4801 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4861 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4921 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4981 taaacttggt ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga
     5041 tcagtgaggc accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc
     5101 ccgtcgtgta gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga
     5161 tgccgcgaga gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga
     5221 gggccgagcg aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct
     5281 gtcgtgatgc tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg
     5341 ctactggcat cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc
     5401 agcggtcaag ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag
     5461 ggcctccgat cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag
     5521 cactacacaa ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt
     5581 actcaaccaa gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt
     5641 ctatacggga caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc
     5701 gttcttcggg gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc
     5761 ccactcttgc acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg
     5821 caaaaacagg caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga
     5881 tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct
     5941 ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata
     6001 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     6061 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     6121 agtgcaggtg ccagaacatt tctct
//
