LOCUS       sequence-175318-        6321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  promoter reporter construct to test 5' flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6321)
  AUTHORS   Sayegh R, Auerbach SD, Li X, Loftus RW, Husted RF, Stokes JB, Thomas
            CP
  TITLE     Glucocorticoid induction of epithelial sodium channel expression in 
            lung and renal epithelia occurs via trans-activation of a hormone 
            response element in the 5'-flanking region of the human epithelial 
            sodium channel alpha subunit gene.
  JOURNAL   J Biol Chem. 1999 Apr 30;274(18):12431-7.
  PUBMED    10212217
REFERENCE   2  (bases 1 to 6321)
  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: "J Biol 
            Chem."; date: "1999-04-30"; volume: "274(18)"; pages: "12431-7"
COMMENT     Sequence Label: hum alpha ENAC promoter a25-a23
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1585..1594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1591..3243
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1657..1675)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3284..3405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3295..3314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3349..3368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3653..3670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3824..4412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3904..3923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4583..5374,5375..5443))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5206..5225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5444..5548)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5575..6030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5662..5681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5872..5893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6161..6209
                     /note="synthetic polyadenylation signal"
     misc_feature    6223..6314
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6263..6282
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg gcttaccaag
       61 cacccagaga gcagacgaat ccacgggctc tgtgtgggag tgagggaggc cttcccggct
      121 ttcacatcca ggtgcacctg agccccgatc ccccatgagt ctgtctcagg aagtaaatgg
      181 caaaagcgct aagtagatag ccccagagga ggaggagaat tctgaattct gtttatagct
      241 ctgctcctat tttgttttct aacctcagaa aactgattta tccttgggaa gggtcagttt
      301 cttcattagg acaatgaaga aaaatccagc tgtcccttcc aaggggaggt atcatgagca
      361 gtatcaaggt aagcaaggga acctggtttg gaatcctggt tgacaccggc tttcacttta
      421 agaagtcgca agagactgcc gcaagagagg aagccacaga ccaggttgga ccctgagcct
      481 ggagtactgc cttttctctt ctttgcctgc taaactcctt gcttgtcaag attcagcaga
      541 gatgacacct tctctgggaa ggcttcctga gtctcctgca gtgagcccct aggctcccct
      601 agcccttggc tcttatgtct tatctctgag aaatctgaga tctgcacttt tgttagaagt
      661 ccgttttaaa gacaatcttt taaagtgtac aaatagaaac cttatcttat tgtattgcag
      721 gcctatctcc tcccccggtg tgtatcaatg ctcaggctgg gcctttgtct gctggcttgt
      781 ggagggaggc aggacaagcc agtgaccaca ttcctgcact ctgggctgcc tcctgtgggg
      841 cccgtgagga agcggcagcc ttccctgccg cgtgcagggc ctgggttgtg tgggtggctc
      901 tcccgagtct cctccagccc tttttgggtc tgattctctg accctcctct ctctccttgc
      961 tcaccacttg gcactccact ccttatcctt ttgctgcatt aagtttggaa agagattttg
     1021 gagaaaatat agcccatcaa cgaatttctc ctcctcccag ggcctgtcta ggcgtgtgcc
     1081 atgcccaccc tcttcctttc cagcgctggc cacatcctcc ctgcaccttc agtgcctgct
     1141 ttccctgcct cttcctgggc tccgggtctg tgtccacagt gtcctgcgtg gccctcgcgc
     1201 tgcctctggt tgcccacatt cctgcaactc tgtgaccaca gcaggggcga ttacacattc
     1261 ctggcctggc agccaacagt gtaaaaaaga acagaatgtc ctagggcccc gcctagcccc
     1321 cagcttcacc tgggcccctc ccgggtctgg acaaggttgg agggggtggc gaggaatcag
     1381 caggaaagag gagggaccag gaggaggcag acgcatcccc ctgagtgcgc tgctcccact
     1441 tagtgagcgg ggaggagacc tgcagagacc tcttctctct tctctgcagg gcctgaaagc
     1501 cgaattctgc agatatccat cacactggcg gccgctcgag atctgcgatc taagtaagct
     1561 tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc
     1621 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
     1681 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
     1741 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
     1801 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
     1861 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     1921 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
     1981 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
     2041 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     2101 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
     2161 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
     2221 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
     2281 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     2341 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     2401 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     2461 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     2521 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     2581 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     2641 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     2701 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     2761 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     2821 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     2881 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     2941 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     3001 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     3061 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     3121 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     3181 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     3241 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     3301 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3361 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3421 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3481 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa
     3541 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     3601 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
     3661 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3721 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     4021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4261 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4381 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4441 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4501 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4561 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4621 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4681 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4741 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4801 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4861 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4921 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4981 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5041 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5101 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5161 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5221 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5281 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5341 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5401 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5461 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5521 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     5581 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5641 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5701 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5761 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5821 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5881 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5941 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6001 attttaacaa aatattaacg tttacaattt cccattcgcc attcaggctg cgcaactgtt
     6061 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag
     6121 taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc
     6181 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     6241 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     6301 cagaacattt ctctatcgat a
//
