LOCUS       sequence-447029-        4550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible middle entry clone containing a destabilized LexA
            transactivator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4550)
  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 4550)
  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 (no LexOp) (JDW 937)
FEATURES             Location/Qualifiers
     source          1..4550
                     /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"
     protein_bind    complement(2437..2536)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2593..2609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2606..2628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2781..3369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2861..2880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3505..4320)
                     /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(3505..4320)
                     /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     4228..4247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4312..4331
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4441..4457
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4441..4457
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4458..4504
                     /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 gccgcgttta aacgctagca actctagatt aattaaaccc agctttcttg tacaaagttg
     2461 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2521 ataaaatcat tatttgaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat
     2581 gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa
     2641 tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg
     2701 gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3181 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc
     3421 gccgtcccgt caagtcagcg taaatgggta gggggcttca aatcgtccgc tctgccagtg
     3481 ttacaaccaa ttaacaaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     3541 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3601 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3661 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3721 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     3781 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3841 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3901 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3961 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     4021 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     4081 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     4141 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     4201 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     4261 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     4321 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     4381 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccgccgc
     4441 tctagaacta gtggatccaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     4501 ttatctgttg tttgtcgcat tatacgagac gtccaggttg ggatacctga
//
