LOCUS       sequence-447027-        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible middle entry clone containing a Destablized LexA 
            and LexOperon with 3' minimal c-Fos promoter for an all in one RU486
            driven gene expression construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  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 5268)
  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-c-Fos (JDW 1359)
FEATURES             Location/Qualifiers
     source          1..5268
                     /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    2404..2525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2441..2460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2495..2514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2544..2635
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2584..2603
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(3155..3254)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3311..3327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3311..3327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3324..3346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3499..4087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3579..3598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4223..5038)
                     /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(4223..5038)
                     /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     4946..4965
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5030..5049
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5159..5175
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5159..5175
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      5176..5222
                     /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 gccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2461 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2521 tcttagatat cgctagcagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     2581 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctcagat
     2641 ctatcgatac gcgtttgcat gccgcttggg ctgcaggtcg aggctaaaaa actaatcgca
     2701 ttatcatccc ctcgacgtac tgtacatata accactggtt ttatatacag cagtactgta
     2761 catataacca ctggttttat atacagcatc tagatactgt acatataacc actggtttta
     2821 tatacagcag tactgtacat ataaccactg gttttatata cagcagaatt cgtactgtac
     2881 atataaccac tggttttata tacagcagta ctgtacatat aaccactggt tttatataca
     2941 gcagtcgagg taagattaga tatggatatg tatatggata tgtatatggt ggtaatgcca
     3001 tgtaatatgc tcgactctag gatcttcgtc gacggatccc cagtgacgta ggaagtccat
     3061 ccattcacag cgcttctata aaggcgccag ctgaggcgcc tactactcca accgcgactg
     3121 cagcgagcaa ctctagcaac tctagattaa ttaaacccag ctttcttgta caaagttggc
     3181 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     3241 aaaatcatta tttgaggcta ggtggaggct cagtgatgat aagtctgcga tggtggatgc
     3301 atgtgtcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaga gggcacaatc
     3361 ctattccgcg ctatccgaca atctccaaga cattaggtgg agttcagttc ggcgtatggc
     3421 atatgtcgct ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3901 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctctattca acaaagccgc
     4141 cgtcccgtca agtcagcgta aatgggtagg gggcttcaaa tcgtccgctc tgccagtgtt
     4201 acaaccaatt aacaaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     4261 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     4321 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     4381 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     4441 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     4501 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     4561 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4621 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4681 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4741 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4801 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     4861 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4921 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4981 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     5041 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     5101 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccgccgctc
     5161 tagaactagt ggatccaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5221 atctgttgtt tgtcgcatta tacgagacgt ccaggttggg atacctga
//
