LOCUS       sequence-414736-        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter construct containing 4 consensus PGR binding 
            sites upstream of a minimal TK promoter, derived from Addgene 
            #11350.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Aarts MT, Wagner M, van der Wal T, van Boxtel AL, van Amerongen R
  TITLE     A molecular toolbox to study progesterone receptor signaling.
  JOURNAL   J Mammary Gland Biol Neoplasia. 2023 Nov 29;28(1):24. doi: 
            10.1007/s10911-023-09550-0.
  PUBMED    38019315
REFERENCE   2  (bases 1 to 6763)
  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 Mammary 
            Gland Biol Neoplasia."; date: "2023-11-29"; volume: "28(1):24. doi";
            pages: " 10.1007/s10911-023-09550-0"
COMMENT     Sequence Label: 4X PRE TK luc
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1976..1994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2154..2173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2655..3168)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2850..2871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3062..3081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3594..3616
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3608..3625
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3609..3625
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3848..3910
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             3970..5622
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4036..4054)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5753..5818
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5948..5968
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6393..6527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6430..6449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6484..6503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6545..6561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6545..6561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6558..6580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6569..6585
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6593..6599,6600..6617,6618..6623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6638..6659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1981 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2041 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2101 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2161 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2221 gagaaaatac cgcatcaggc gattccaaca tccaataaat catacaggca aggcaaagaa
     2281 ttagcaaaat taagcaataa agcctcagag cataaagcta aatcggttgt accaaaaaca
     2341 ttatgaccct gtaatacttt tgcgggagaa gcctttattt caacgcaagg ataaaaattt
     2401 ttagaaccct catatatttt aaatgcaatg cctgagtaat gtgtaggtaa agattcaaac
     2461 gggtgagaaa ggccggagac agtcaaatca ccatcaatat gatattcaac cgttctagct
     2521 gataaattca tgccggagag ggtagctatt tttgagaggt ctctacaaag gctatcaggt
     2581 cattgcctga gagtctggag caaacaagag aatcgatgaa cggtaatcgt aaaactagca
     2641 tgtcaatcat atgtaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     2701 aagcaaatat ttaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3121 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     3181 ttgacgagca cgtataacgt gctttcctcg ttagaatcag agcgggagct aaacaggagg
     3241 ccgattaaag ggattttaga caggaacggt acgccagaat cctgagaagt gtttttataa
     3301 tcagtgaggc caccgagtaa aagagtctgt ccatcacgca aattaaccgt tgtcgcaata
     3361 cttctttgat tagtaataac atcacttgcc tgagtagaag aactcaaact atcggccttg
     3421 ctggtaatat ccagaacaat attaccgcca gccattgcaa cggaatcgcc attcgccatt
     3481 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3541 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3601 acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg actctagagg
     3661 atctgtagct agaacatcct gtacagtatc cgtagctaga acatcctgta caggatctgt
     3721 agctagaaca tcctgtacag tatccgtagc tagaacatcc tgtacaggat ccggccccgc
     3781 ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccga
     3841 ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg
     3901 acccgcttaa cagcgtcaac agcgtgccgc agatctcgag gagcttggca ttccggtact
     3961 gttggtaaaa tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta
     4021 gaggatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct
     4081 ggaacaattg cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc
     4141 gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga
     4201 atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt
     4261 atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt
     4321 atgaacattt cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg
     4381 aacgtgcaaa aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat
     4441 taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat
     4501 gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat
     4561 tcctctggat ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc
     4621 agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt
     4681 ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata
     4741 tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt
     4801 caggattaca aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa
     4861 agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca
     4921 cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga
     4981 caaggatatg ggctcactga gactacatca gctattctga ttacacccga gggggatgat
     5041 aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat
     5101 accgggaaaa cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt
     5161 atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg
     5221 ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc
     5281 ttgaagtctt taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata
     5341 ttgttacaac accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc
     5401 ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag
     5461 atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg
     5521 tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag
     5581 atcctcataa aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc
     5641 gatgacgaaa ttcttagcta ttgtaatact ctagaggatc tttgtgaagg aaccttactt
     5701 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
     5761 aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat
     5821 tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct
     5881 gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc
     5941 tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct
     6001 aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac
     6061 cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt
     6121 tataagtagg cataacagtt ataatcataa catactgttt tttcttactc cacacaggca
     6181 tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg
     6241 taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc
     6301 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     6361 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     6421 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6481 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattcgtaat
     6541 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6601 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6661 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6721 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcg
//
