LOCUS       sequence-447028-        5409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible middle entry clone containing a Destablized LexA 
            and LexOperon with minimal CaMV 35S promoter as an all in one 
            casette for RU486 driven gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5409)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 5409)
  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: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pME-LexA-mODC-LexOP-35SCaMV (JDW 1348)
FEATURES             Location/Qualifiers
     source          1..5409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..90
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     82..99
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     83..99
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    152..251
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      270..279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             300..551
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     CDS             1866..2237
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    2506..2627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2543..2562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2597..2616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3197..3243
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     protein_bind    complement(3296..3395)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3452..3468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3452..3468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3465..3487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3640..4228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3720..3739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4364..5179)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4364..5179)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5087..5106
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5171..5190
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5300..5316
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5300..5316
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      5317..5363
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aacaaaaccc atcgtacggc caaggaagtc tccaataact gtgatccacc acaagcgcca
       61 gggttttccc agtcacgacg ttgtaaaacg acggccagtc atgcataatc cgcacgcatc
      121 tggaataagg aagtgccatt ccgcctgacc tcaaataatg attttatttt gactgatagt
      181 gacctgttcg ttgcaacaga ttgatgagca atgctttttt ataatgccaa ctttgtacaa
      241 aaaagcaggc ttacgtaact agtggatccg ccaccatggc acccaagaag aagaggaaga
      301 tgaaagcgtt aacggccagg caacaagagg tgtttgatct catccgtgat cacatcagcc
      361 agacaggtat gccgccgacg cgtgcggaaa tcgcgcagcg tttggggttc cgttccccaa
      421 acgcggctga agaacatctg aaggcgctgg cacgcaaagg cgttattgaa attgtttccg
      481 gcgcatcacg cgggattcgt ctgttgcagg aagaggaaga agggttgccg ctggtaggtc
      541 gtgtggctgc cggtgaaccg tcgagcgccg gtaccgaatt cccgggtgtc gaccagaaaa
      601 agttcaataa agtcagagtt gtgagagcac tggatgctgt tgctctccca cagccagtgg
      661 gcgttccaaa tgaaagccaa gccctaagcc agagattcac tttttcacca ggtcaagaca
      721 tacagttgat tccaccactg atcaacctgt taatgagcat tgaaccagat gtgatctatg
      781 caggacatga caacacaaaa cctgacacct ccagttcttt gctgacaagt cttaatcaac
      841 taggcgagag gcaacttctt tcagtagtca agtggtctaa atcattgcca ggttttcgaa
      901 acttacatat tgatgaccag ataactctca ttcagtattc ttggatgagc ttaatggtgt
      961 ttggtctagg atggagatcc tacaaacacg tcagtgggca gatgctgtat tttgcacctg
     1021 atctaatact aaatgaacag cggatgaaag aatcatcatt ctattcatta tgccttacca
     1081 tgtggcagat cccacaggag tttgtcaagc ttcaagttag ccaagaagag ttcctctgta
     1141 tgaaagtatt gttacttctt aatacaattc ctttggaagg gctacgaagt caaacccagt
     1201 ttgaggagat gaggtcaagc tacattagag agctcatcaa ggcaattggt ttgaggcaaa
     1261 aaggagttgt gtcgagctca cagcgtttct atcaacttac aaaacttctt gataacttgc
     1321 atgatcttgt caaacaactt catctgtact gcttgaatac atttatccag tcccgggcac
     1381 tgagtgttga atttccagaa atgatgtctg aagttattgc tgggtcgacg cccatggaat
     1441 ttcagtacct gccagataca gacgatcgtc accggattga ggagaaacgt aaaaggacat
     1501 atgagacctt caagagcatc atgaagaaga gtcctttcag cggacccacc gacccccggc
     1561 ctccacctcg acgcattgct gtgccttccc gcagctcagc ttctgtcccc aagccagcac
     1621 cccagcccta tccctttacg tcatccctga gcaccatcaa ctatgatgag tttcccacca
     1681 tggtgtttcc ttctgggcag atcagccagg cctcggcctt ggccccggcc cctccccaag
     1741 tcctgcccca ggctccagcc cctgcccctg ctccagccat ggtatcagct ctggcccagg
     1801 ccccagcccc tgtcccagtc ctagccccag gccctcctca ggctgtggcc ccacctgccc
     1861 ccaagcccac ccaggctggg gaaggaacgc tgtcagaggc cctgctgcag ctgcagtttg
     1921 atgatgaaga cctgggggcc ttgcttggca acagcacaga cccagctgtg ttcacagacc
     1981 tggcatccgt cgacaactcc gagtttcagc agctgctgaa ccagggcata cctgtggccc
     2041 cccacacaac tgagcccatg ctgatggagt accctgaggc tataactcgc ctagtgacag
     2101 gggcccagag gccccccgac ccagctcctg ctccactggg ggccccgggg ctccccaatg
     2161 gcctcctttc aggagatgaa gacttctcct ccattgcgga catggacttc tcagccctgc
     2221 tgagtcagat cagctccaga tctcgagata tcagccatgg cttcccgccg gcggtggcgg
     2281 cgcaggatga tggcacgctg cccatgtctt gtgcccagga gagcgggatg gaccgtcacc
     2341 ctgcagcctg tgcttctgct aggatcaatg tgtagtaaat cgatgatcct ccacgtagcg
     2401 gccgcaatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2461 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagt ttaaacgcgt
     2641 taacaattgg aaattaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     2701 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     2761 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     2821 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga
     2881 attgatcccc cctcgacagc ttgcatgccg cttgggctgc aggtcgaggc taaaaaacta
     2941 atcgcattat catcccctcg acgtactgta catataacca ctggttttat atacagcagt
     3001 actgtacata taaccactgg ttttatatac agcagtcgac gtactgtaca tataaccact
     3061 ggttttatat acagcagtac tgtacatata accactggtt ttatatacag cagtcgaggt
     3121 aagattagat atggatatgt atatggatat gtatatggtg gtaatgccat gtaatatgct
     3181 cgactctagg atcttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     3241 ggacacgctg aagctagtcg actctagcct cggctagcaa ctctagatta attaaaccca
     3301 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     3361 caggtcacta tcagtcaaaa taaaatcatt atttgaggct aggtggaggc tcagtgatga
     3421 taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt
     3481 atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg
     3541 gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca
     3601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4261 cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa
     4321 atcgtccgct ctgccagtgt tacaaccaat taacaaattc tgattagaaa aactcatcga
     4381 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     4441 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     4501 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     4561 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     4621 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     4681 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     4741 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     4801 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     4861 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4921 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4981 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     5041 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     5101 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     5161 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     5221 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     5281 cgtggctttc ccccgccgct ctagaactag tggatccaaa taaaacgaaa ggctcagtcg
     5341 aaagactggg cctttcgttt tatctgttgt ttgtcgcatt atacgagacg tccaggttgg
     5401 gatacctga
//
