LOCUS       sequence-414738-        6871 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Luciferase reporter construct containing 8 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 6871)
  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 6871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: 8X PRE TK luc
FEATURES             Location/Qualifiers
     source          1..6871
                     /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        3956..4018
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             4078..5730
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4144..4162)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5861..5926
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6056..6076
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6501..6635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6538..6557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6592..6611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6653..6669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6653..6669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6666..6688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6677..6693
                     /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(6701..6707,6708..6725,6726..6731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6746..6767
                     /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 tacaggatcc ggccccgccc agcgtcttgt
     3901 cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtccgagg tccacttcgc
     3961 atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaca
     4021 gcgtcaacag cgtgccgcag atctcgagga gcttggcatt ccggtactgt tggtaaaatg
     4081 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atcctctaga ggatggaacc
     4141 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     4201 tttacagatg cacatatcga ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt
     4261 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     4321 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     4381 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gaacatttcg
     4441 cagcctaccg tagtgtttgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     4501 aaattaccaa taatccagaa aattattatc atggattcta aaacggatta ccagggattt
     4561 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     4621 gtaccagagt cctttgatcg tgacaaaaca attgcactga taatgaattc ctctggatct
     4681 actgggttac ctaagggtgt ggcccttccg catagaactg cctgcgtcag attctcgcat
     4741 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     4801 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     4861 gtcgtcttaa tgtatagatt tgaagaagag ctgtttttac gatcccttca ggattacaaa
     4921 attcaaagtg cgttgctagt accaacccta ttttcattct tcgccaaaag cactctgatt
     4981 gacaaatacg atttatctaa tttacacgaa attgcttctg ggggcgcacc tctttcgaaa
     5041 gaagtcgggg aagcggttgc aaaacgcttc catcttccag ggatacgaca aggatatggg
     5101 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     5161 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     5221 ctgggcgtta atcagagagg cgaattatgt gtcagaggac ctatgattat gtccggttat
     5281 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     5341 gacatagctt actgggacga agacgaacac ttcttcatag ttgaccgctt gaagtcttta
     5401 attaaataca aaggatatca ggtggccccc gctgaattgg aatcgatatt gttacaacac
     5461 cccaacatct tcgacgcggg cgtggcaggt cttcccgacg atgacgccgg tgaacttccc
     5521 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     5581 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     5641 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     5701 gccaagaagg gcggaaagtc caaattgtaa aatgtaactg tattcagcga tgacgaaatt
     5761 cttagctatt gtaatactct agaggatctt tgtgaaggaa ccttacttct gtggtgtgac
     5821 ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag
     5881 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg
     5941 aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga
     6001 agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa
     6061 aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag
     6121 tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa
     6181 agctgcactg ctatacaaga aaattatgga aaaatattct gtaaccttta taagtaggca
     6241 taacagttat aatcataaca tactgttttt tcttactcca cacaggcata gagtgtctgc
     6301 tattaataac tatgctcaaa aattgtgtac ctttagcttt ttaatttgta aaggggttaa
     6361 taaggaatat ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg
     6421 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     6481 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     6541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6601 ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttcgtaatca tggtcatagc
     6661 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6721 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6841 gcgcggggag aggcggtttg cgtattgggc g
//
