LOCUS       sequence-414742-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMuLE R4-R3 entry plasmid containing human full length PRB gene, for
            3-component Gateway assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  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 6533)
  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: pMuLE (R4-R3) PRB
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             4515..4526
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4596..4719
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4774..4793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4775..4793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4798..4814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4798..4814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4916..4935)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4927..5736
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4994..5013)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5883..6471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6372..6391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgggag acccaagctt ggtaccatga ctgagctgaa ggcaaagggt ccccgggctc
      781 cccacgtggc gggcggcccg ccctcccccg aggtcggatc cccactgctg tgtcgcccag
      841 ccgcaggtcc gttcccgggg agccagacct cggacacctt gcctgaagtt tcggccatac
      901 ctatctccct ggacgggcta ctcttccctc ggccctgcca gggacaggac ccctccgacg
      961 aaaagacgca ggaccagcag tcgctgtcgg acgtggaggg cgcatattcc agagctgaag
     1021 ctacaagggg tgctggaggc agcagttcta gtcccccaga aaaggacagc ggactgctgg
     1081 acagtgtctt ggacactctg ttggcgccct caggtcccgg gcagagccaa cccagccctc
     1141 ccgcctgcga ggtcaccagc tcttggtgcc tgtttggccc cgaacttccc gaagatccac
     1201 cggctgcccc cgccacccag cgggtgttgt ccccgctcat gagccggtcc gggtgcaagg
     1261 ttggagacag ctccgggacg gcagctgccc ataaagtgct gccccggggc ctgtcaccag
     1321 cccggcagct gctgctcccg gcctctgaga gccctcactg gtccggggcc ccagtgaagc
     1381 cgtctccgca ggccgctgcg gtggaggttg aggaggagga tggctctgag tccgaggagt
     1441 ctgcgggtcc gcttctgaag ggcaaacctc gggctctggg tggcgcggcg gctggaggag
     1501 gagccgcggc tgtcccgccg ggggcggcag caggaggcgt cgccctggtc cccaaggaag
     1561 attcccgctt ctcagcgccc agggtcgccc tggtggagca ggacgcgccg atggcgcccg
     1621 ggcgctcccc gctggccacc acggtgatgg atttcatcca cgtgcctatc ctgcctctca
     1681 atcacgcctt attggcagcc cgcactcggc agctgctgga agacgaaagt tacgacggcg
     1741 gggccggggc tgccagcgcc tttgccccgc cgcggagttc accctgtgcc tcgtccaccc
     1801 cggtcgctgt aggcgacttc cccgactgcg cgtacccgcc cgacgccgag cccaaggacg
     1861 acgcgtaccc tctctatagc gacttccagc cgcccgctct aaagataaag gaggaggagg
     1921 aaggtgcgga ggcctccgcg cgctccccgc gttcctacct tgtggccggt gccaaccccg
     1981 cagccttccc ggatttcccg ttggggccac cgcccccgct gccgccgcga gcgaccccat
     2041 ccagacccgg ggaagcggcg gtgacggccg cacccgccag tgcctcagtc tcgtctgcgt
     2101 cctcctcggg gtcgaccctg gagtgcatcc tgtacaaagc ggagggcgcg ccgccccagc
     2161 agggcccgtt cgcgccgccg ccctgcaagg cgccgggcgc gagcggctgc ctgctcccgc
     2221 gggacggcct gccctccacc tccgcctctg ccgccgccgc cggggcggcc cccgcgctct
     2281 accctgcact cggcctcaac gggctcccgc agctcggcta ccaggccgcc gtgctcaagg
     2341 agggcctgcc gcaggtctac ccgccctatc tcaactacct gaggccggat tcagaagcca
     2401 gccagagccc acaatacagc ttcgagtcat tacctcagaa gatttgttta atctgtgggg
     2461 atgaagcatc aggctgtcat tatggtgtcc ttacctgtgg gagctgtaag gtcttcttta
     2521 agagggcaat ggaagggcag cacaactact tatgtgctgg aagaaatgac tgcatcgttg
     2581 ataaaatccg cagaaaaaac tgcccagcat gtcgccttag aaagtgctgt caggctggca
     2641 tggtccttgg aggtcgaaaa tttaaaaagt tcaataaagt cagagttgtg agagcactgg
     2701 atgctgttgc tctcccacag ccattgggcg ttccaaatga aagccaagcc ctaagccaga
     2761 gattcacttt ttcaccaggt caagacatac agttgattcc accactgatc aacctgttaa
     2821 tgagcattga accagatgtg atctatgcag gacatgacaa cacaaaacct gacacctcca
     2881 gttctttgct gacaagtctt aatcaactag gcgagaggca acttctttca gtagtcaagt
     2941 ggtctaaatc attgccaggt tttcgaaact tacatattga tgaccagata actctcattc
     3001 agtattcttg gatgagctta atggtgtttg gtctaggatg gagatcctac aaacatgtca
     3061 gtgggcagat gctgtatttt gcacctgatc taatactaaa tgaacagcgg atgaaagaat
     3121 catcattcta ttcattatgc cttaccatgt ggcagatccc acaggagttt gtcaagcttc
     3181 aagttagcca agaagagttc ctctgtatga aagtattgtt acttcttaat acaattcctt
     3241 tggaagggct acgaagtcaa acccagtttg aggagatgag gtcaagctac attagagagc
     3301 tcatcaaggc aattggtttg aggcaaaaag gagttgtgtc gagctcacag cgtttctatc
     3361 aacttacaaa acttcttgat aacttgcatg atcttgtcaa acagcttcat ctgtactgct
     3421 tgaatacatt tatccagtcc cgggcactga gtgttgaatt tccagaaatg atgtctgaag
     3481 ttattgctgc acaattaccc aagatattgg cagggatggt gaaacccctt ctctttcata
     3541 aaaagtgaat gtcatctttt tcttttaaag aattaaattt tgtggcatgt ctttttgttt
     3601 tggtcaggat tatgaggtct tgagttttta taatgttctt ctgaaagcct tacatttata
     3661 acatcatagt gtgtaaattt aaaagaaaaa ttgtgaggtt ctaattattt tcttttataa
     3721 agtataatta gaatgtttaa ctgttttgtt tacccatatt ttcttgaaga atttacaaga
     3781 ttgaaaaagt actaaaattg ttaaagtaaa ctatatctta tccatattat ttcataccat
     3841 gtaggtgagg atttttaact tttgcatcta acaaatcatc gacttaagag aaaaaatctt
     3901 acatgtaata acacaaagct attatatgtt atttctaggt aactcccttt gtgtcaatta
     3961 tatttccaaa aatgaacctt taaaatggta tgcaaaattt tgtctatata tatttgtgtg
     4021 aggaggaaat tcataacttt cctcagattt tcaaaagtat ttttaatgca aaaaatgtag
     4081 aaagagttta aaaccactaa aatagattga tgttcttcaa actaggcaaa acaactcata
     4141 tgttaagacc attttccaga ttggaaacac aaatctctta ggaagttaat aagtagattc
     4201 atatcattat acaaatagta ttgtgggttt tgtaggtttt taaaataacc ttttttgggg
     4261 agagaattgt cctctaatga ggtattgcga gtggacataa gaaatcagaa gattatggcc
     4321 taactgtact ccttaccaac tgtggcatgc tgaaagttag tcactcttac tgattctcaa
     4381 ttctctcacc tttgaaagta gtaaaatatc tttcctgcca attgctcctt tgggtcagag
     4441 cttattaaca tcttttcaaa tcaaaggaaa gaagaaaggg agaggaggag gagggagata
     4501 tcaattcaca tacctttctc ctctttatcc tccactatca tgaattctgc agatatccat
     4561 cacactggcg gccgctcgag catgcatcta gaggacaact ttgtataata aagttgaacg
     4621 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
     4681 tacataatac tgtaaaacac aacatatcca gtcactatga atcaactact tagatggtat
     4741 tagtgacctg tactgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc
     4801 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     4861 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     4921 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     4981 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     5041 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     5101 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     5161 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     5221 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     5281 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     5341 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     5401 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     5461 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     5521 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     5581 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     5641 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     5701 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     5761 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     5821 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
