LOCUS       sequence-175317-        8142 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 8142)
  AUTHORS   Itani OA, Liu KZ, Cornish KL, Campbell JR, Thomas CP
  TITLE     Glucocorticoids stimulate human sgk1 gene expression by activation 
            of a GRE in its 5'-flanking region.
  JOURNAL   Am J Physiol Endocrinol Metab. 2002 Nov;283(5):E971-9.
  PUBMED    12376324
REFERENCE   2  (bases 1 to 8142)
  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: "Am J 
            Physiol Endocrinol Metab."; date: "2002-11"; volume: "283(5)"; 
            pages: "E971-9"
COMMENT     Sequence Label: hum sgk promoter F4R4
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3406..3415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3412..5064
                     /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(3478..3496)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5105..5226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5170..5189
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5474..5491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5645..6233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5725..5744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6404..7195,7196..7264))
                     /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     7027..7046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7265..7369)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7396..7851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7483..7502)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7693..7714
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7982..8030
                     /note="synthetic polyadenylation signal"
     misc_feature    8044..8135
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     8084..8103
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccctttgcgt
       61 gctgggggtg taataaagct aggacactga tgactgtaat tttctgagtg atgtcgcaat
      121 ggggagaata aatggcccag tgttgaactt tgcagcttgc tcccttcctg tacttaaatt
      181 gccttgatat aaagtggggt tcataacaga acagggatag ccgtctctgg ctcgtgctct
      241 catgtcatct cagagttcca gcttatcaga ggcatgtagc agggaggctt attccagcca
      301 taactgggct ctacctccag cctccagaag taatccccaa cctgcatatc cttgggcaac
      361 ccgaagaatg aaagaagaag ctgtaaaacc ccctttgaaa ggttcgtact taccgtacta
      421 tattttgcag atgcctcaaa ggatttgggg ttacttggca tggggaaggc acataaggtg
      481 gggtgtagga gagggtctct ggttgtaggt ttcttaattt aatgtttgaa aacaaacatg
      541 caaaagtctg tgtgcaggtt gatgtttctg ggcagcctga gcaaaatttg ctctctcaag
      601 agggaaagga accaggtggg agcagagcta ggctgggcta ggctagttga atggtgggac
      661 atgacatacg ggtggcactg gcaataacaa agtcacattc tatgaagatt ccctgcaaga
      721 ggaagcagac atgggccagt tactgtgatt tgaaattgcc taaacattgc tttaggttgg
      781 catgtcaatt tcaggtacta gtgttttttt tgtttttgtt tttttttgtt tttgtttgtt
      841 tgtttgtttt gagacggagt ctcgctctgt tgccaggctg gagtgcagtg gcgtgatctc
      901 ggctcactgc aacctccgcc tcccgggttc aagcgattct cctgcctcag cctcccgagt
      961 agctgggact acaggcgcac gccaccacgc ctggctaact tttctatttt cagtagagac
     1021 ggggtttcac catgttagcc aggatggtct cgatctcttg acctcgtgat ccgcccgcct
     1081 tggcctccca aagtgctggg attacaggcg tgagcccctg cgcccggccc cagtaaatgc
     1141 tttttataag tgtgggcact gagcaaactt tcccagccag actccaggag agagaatgtg
     1201 tttcccttct ctcggtttgg ggctgttgca acaaagcaaa ccaaggagtt gagactagag
     1261 ctcactttag ggcaagtggg ggtggttttg cctgcaaaac aaacccctgc ccaagaccaa
     1321 ggaaaaggcg tttcacatgc tattcctggt ttgacagctg gtatttcggg actgtgccag
     1381 atccagtagg caactttaaa atggcagagc ctttggtagc aagaggtcat ggcagggcag
     1441 ccaccgcaga cagcaacagc gagcgccagg tacctggccc tgcgaatagt ggtaacttgt
     1501 aactgcccgc tccgggccca gtcgctgtgc tcgcggcttc ccggccagca ctggctcacg
     1561 tccccgcgcc ggcggtcagg ctgcggctcc cagacatccc ccagccgcgg ggttactgga
     1621 aggcaccggc atcgctgttc tgcagagccc gggccgccgc ctcgagcttc cctctcttcc
     1681 ctgccttctg cagcggagtc acccggctaa tctttcagga taaagtcaca gtttatgtgg
     1741 gactcacata aagagcgagc gaggtggcaa aactaagaag ccctggggca gccttgagtt
     1801 aaacccaggg agggtaggga cgattttaag accatgtatc atgacctgca gggttttcag
     1861 gtgggacagc gggagaggag caggccccac agaggaatcg aggatgcccg gttcacgcca
     1921 ggtctgcccc cgggcaaagc tacccctccc ttcgcttgtt acctcctcac gtgttcttgg
     1981 catggcagag attaaaaatg caaggaaaaa aattacatgc ggaacggaca aaatgttctc
     2041 agagattact tcagaaaaaa aaaagtgaaa tgcagattgt acttcttcct ttagtgcaga
     2101 gacgactttt atttccgccc cctcccctcc acattcctga cctctccctc ccccttttcc
     2161 ctctttcttt ccttccttcc tcctcttcca agttctggga tttttcagcc ttgcttggtt
     2221 ttggccaaaa gcacaaaaaa ggcgttttcg gaagcgaccc gaccgtgcac aagggccatt
     2281 tgtttgtttt gggactcggg gcaggaaatc ttgcccggcc tgagtcacgg cggctccttc
     2341 aaggaaacgt cagtgctcgc cggtcgctct cgtctgccgc gcgccccgcc gcccgctgcc
     2401 catgggggag atgcagggcg cgctggccag agcccggctc gagtccctgc tgcggccccg
     2461 ccacaaaaag agggccgagg cgcagaaaag gagcgagtcc ttcctgctga gcggactggg
     2521 taagcgccgc cgccggcccc gctgggggct tggctcactt ccccagagcg gcttggaggc
     2581 aggggccggc tttcgtcgga gttctcgggg ccggggtccc ggcggcggga acgggaggac
     2641 ctggcgggcg aggtcgcgcg cgcaggcccg cgccccaggg ataaaccccg gagggtggcg
     2701 cgcaccgccg gctcgggttg gggaggaggg tgggagtccg gccgcaggac ggcgcctggc
     2761 cggggagagg gtatctgcag ggacagtgag cgaagccacc gtggccgccg cgcacccgcc
     2821 gggaagcgct tcggcgctgc gaacccggct ttctccggcg gcggaataaa tgagagaggt
     2881 ggaaaactac cccgggctct ccggccctcc ccgcgccctc cgccggcgcg ttctctctct
     2941 cctgccccag gagccgatgg agactgacaa cggccctgcg ccaggccgtc cccgggcggt
     3001 cctcgcgccc ccgcccgggg ctcgccctct caatggggac agaaccgccc gccgcaggca
     3061 gcgtagccgc cagcaaaccg cgaggcggtc ggggcggggc gaggggcgag gcgaagggcg
     3121 gggccacttc tcactgtcgc gcaggccccg cccccgcggc ggtgcctttt ttataaggcc
     3181 gagcgcgcgg cctggcgcag catacgccga gccggtcttt gagcgctaac gtctttctgt
     3241 ctccccgcgg tggtgatgac ggtgaaaact gaggctgcta agggcaccct cacttactcc
     3301 aggatgaggg gcatggaagg gcgaattctg cagatatcca tcacactggc ggccgctcga
     3361 gatctgcgat ctaagtaagc ttggcattcc ggtactgttg gtaaagccac catggaagac
     3421 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     3481 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     3541 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     3601 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     3661 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     3721 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     3781 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     3841 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     3901 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     3961 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     4021 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     4081 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     4141 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     4201 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     4261 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     4321 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     4381 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     4441 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     4501 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     4561 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     4621 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     4681 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     4741 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     4801 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     4861 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     4921 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     4981 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     5041 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     5101 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5161 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5221 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     5281 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     5341 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     5401 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     5461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5821 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     5881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6061 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7381 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7441 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7501 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7561 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7621 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7681 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7741 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7801 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcccattcgc
     7861 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7921 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     7981 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     8041 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     8101 ccccagtgca agtgcaggtg ccagaacatt tctctatcga ta
//
