LOCUS       sequence-414740-        6979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter construct containing 12 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 6979)
  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 6979)
  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: 12x PRE TK luc
FEATURES             Location/Qualifiers
     source          1..6979
                     /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        4064..4126
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             4186..5838
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4252..4270)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5969..6034
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6164..6184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6609..6743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6646..6665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6700..6719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6761..6777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6761..6777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6774..6796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6785..6801
                     /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(6809..6815,6816..6833,6834..6839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6854..6875
                     /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 ctgtagctag
     3781 aacatcctgt acagtatccg tagctagaac atcctgtaca ggatctgtag ctagaacatc
     3841 ctgtacagta tccgtagcta gaacatcctg tacaggatct gtagctagaa catcctgtac
     3901 agtatccgta gctagaacat cctgtacagg atctgtagct agaacatcct gtacagtatc
     3961 cgtagctaga acatcctgta caggatccgg ccccgcccag cgtcttgtca ttggcgaatt
     4021 cgaacacgca gatgcagtcg gggcggcgcg gtccgaggtc cacttcgcat attaaggtga
     4081 cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg
     4141 tgccgcagat ctcgaggagc ttggcattcc ggtactgttg gtaaaatgga agacgccaaa
     4201 aacataaaga aaggcccggc gccattctat cctctagagg atggaaccgc tggagagcaa
     4261 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
     4321 catatcgagg tgaacatcac gtacgcggaa tacttcgaaa tgtccgttcg gttggcagaa
     4381 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     4441 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     4501 aacgacattt ataatgaacg tgaattgctc aacagtatga acatttcgca gcctaccgta
     4561 gtgtttgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa attaccaata
     4621 atccagaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     4681 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt accagagtcc
     4741 tttgatcgtg acaaaacaat tgcactgata atgaattcct ctggatctac tgggttacct
     4801 aagggtgtgg cccttccgca tagaactgcc tgcgtcagat tctcgcatgc cagagatcct
     4861 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     4921 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     4981 tatagatttg aagaagagct gtttttacga tcccttcagg attacaaaat tcaaagtgcg
     5041 ttgctagtac caaccctatt ttcattcttc gccaaaagca ctctgattga caaatacgat
     5101 ttatctaatt tacacgaaat tgcttctggg ggcgcacctc tttcgaaaga agtcggggaa
     5161 gcggttgcaa aacgcttcca tcttccaggg atacgacaag gatatgggct cactgagact
     5221 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     5281 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     5341 cagagaggcg aattatgtgt cagaggacct atgattatgt ccggttatgt aaacaatccg
     5401 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     5461 tgggacgaag acgaacactt cttcatagtt gaccgcttga agtctttaat taaatacaaa
     5521 ggatatcagg tggcccccgc tgaattggaa tcgatattgt tacaacaccc caacatcttc
     5581 gacgcgggcg tggcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     5641 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     5701 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     5761 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     5821 ggaaagtcca aattgtaaaa tgtaactgta ttcagcgatg acgaaattct tagctattgt
     5881 aatactctag aggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa
     5941 actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt
     6001 taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg
     6061 ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc
     6121 tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa
     6181 ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt
     6241 tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct
     6301 atacaagaaa attatggaaa aatattctgt aacctttata agtaggcata acagttataa
     6361 tcataacata ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta
     6421 tgctcaaaaa ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt
     6481 gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac
     6541 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     6601 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6661 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6721 atgtatctta tcatgtctgg atcgggaatt cgtaatcatg gtcatagctg tttcctgtgt
     6781 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6841 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6901 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6961 gcggtttgcg tattgggcg
//
