LOCUS       sequence-446945-        3974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible middle entry clone containing TAEL2.0 
            transcription factor for making binary TAEL system vectors 
            (generating a tissue specific driver).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3974)
  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 3974)
  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-TAEL (JDW 1324)
FEATURES             Location/Qualifiers
     source          1..3974
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1439..1486
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    1516..1637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1527..1546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1581..1600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    1711..1802
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1751..1770
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(2099..2198)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2215..2234)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2216..2234)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2239..2255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2239..2255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2357..2376)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2368..3177
                     /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(2435..2454)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3324..3912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3813..3832
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg atccactagt ttaattaagc cgccaccatg ttgcctgcag atgcacttga
      721 tgactttgac ctggacatgc tcccagctga tgctctggat gactatgacc tcgacatcct
      781 gcctgggaat gagtttggag cggacgacac tcgtgttgaa gtgcagccac ctgcccagtg
      841 ggtgctcgat ctcatagaag caagtccgat tgcatctgtg gtgtccgacc ctcgcttagc
      901 agacaatcca cttattgcca tcaaccaggc cttcacagat ttaaccggct acagtgagga
      961 agagtgtgtg ggtaggaact gccgctttct ggccgggagc ggaacagagc cctggctgac
     1021 tgacaagatc aggcaaggag tcagagagca taaaccagta ttggttgaga tcctcaacta
     1081 caagaaagat ggaacgccat tcagaaatgc tgttctagta gctcccatct atgatgatga
     1141 tgatgagctg ctgtacttcc tgggctctca ggtggaggtg gacgatgacc agcccaacat
     1201 gggcatggcc agacgagaac gagccgcaga gatgctgaag acactatcgc cgcggcagct
     1261 ggaggtcacc acccttgtgg ctagcgggtt gcgcaacaaa gaagttgcag cgcggttggg
     1321 tctttcagaa aagactgtca agatgcacag aggtctcgtc atggagaaat tgaatctgaa
     1381 aacatctgct gatctggtca ggatagctgt ggaagccggc attgagttct ccgcgtcaaa
     1441 gagacctgct gccactaaaa aagctggaca agccaagaag aaaaaatgat aaccatggaa
     1501 ttctagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1561 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1621 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1681 aggtgtggga ggttttttat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     1741 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     1801 ctcctcgaga gatctttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     1861 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     1921 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     1981 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     2041 gaattgatcc cccctcgaca gcggtaccac gcgtacgcgt gaattcgata tcaagcttac
     2101 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2161 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2221 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2281 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2341 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2401 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2461 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2521 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2581 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2641 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2701 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2761 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2821 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2881 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2941 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     3001 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     3061 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     3121 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     3181 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3241 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3301 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3361 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3421 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3481 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3541 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3601 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3661 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3721 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3781 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3841 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3901 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3961 tttgctcaca tgtt
//
