LOCUS       sequence-486848-        3663 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Gateway middle entry clone containing Dre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3663)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, 
            Largoza G, Sherman M, 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 with diverse applications in vertebrate model 
            systems.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267.
  PUBMED    39026881
REFERENCE   2  (bases 1 to 3663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267."
COMMENT     Sequence Label: pME-DRE-nls (JDW 31)
FEATURES             Location/Qualifiers
     source          1..3663
                     /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"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1745..1765
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1788..1887)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1904..1923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1905..1923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1928..1944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1928..1944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2046..2065)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2057..2866
                     /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(2124..2143)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3013..3601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3502..3521
                     /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 agcaggctcc gcggccgccc ccttcaccgt cgccaccatg ggtgctagcg agctgatcat
      721 ctctggctcc tctggaggat tcctgaggaa catcggcaag gagtaccagg aggctgctga
      781 gaacttcatg agattcatga atgaccaggg agcctacgcc cctaacaccc tgagagacct
      841 gaggctggtg ttccactcct gggctagatg gtgccacgct agacagctgg cctggttccc
      901 tatctctcct gagatggcta gggagtactt ccttcagctg cacgatgctg acctggcctc
      961 taccaccatc gacaagcact acgccatgct gaacatgctg ctgtcccact gtggcctgcc
     1021 tcctctgtct gatgacaagt ctgtgagcct ggccatgagg agaatccgga gagaggctgc
     1081 caccgagaag ggagagagaa ccggccaggc catccctctg agatgggatg acctgaagct
     1141 gctggatgtg ctgctgtcta gatctgagag actggtggac ctgaggaata gggccttcct
     1201 gtttgtggcc tacaacaccc tgatgaggat gtctgagatc tctaggatca gagtgggaga
     1261 cctggaccag accggagaca ccgtgaccct gcacatctcc cacaccaaga ccatcaccac
     1321 cgctgctggc ctggacaaag tgctgtctag gaggaccacc gctgtgctga atgactggct
     1381 ggatgtgtct ggcctgagag agcaccctga cgctgtgctg ttccctccta tccaccggag
     1441 caacaaggct aggatcacca ccacccctct gaccgcccct gccatggaga agatttttag
     1501 cgatgcctgg gtgctgctga acaagaggga tgccacccct aacaagggcc gctaccggac
     1561 ctggaccggc cactctgcta gagtgggagc tgccatcgac atggctgaga agcaagtgtc
     1621 catggtggag atcatgcagg agggcacctg gaaaaagcct gagacactga tgagatacct
     1681 gaggagggga ggagtgtctg tgggagccaa ctctaggctg atggactccg ctagcggcgc
     1741 cggtcctaag aagaagagga aagtgtgatg aaagggtggg cgcgccgacc cagctttctt
     1801 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1861 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     1921 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1981 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2041 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2101 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2161 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2221 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2281 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2341 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2401 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2461 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2521 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2581 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2641 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2701 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2761 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2821 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2881 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     2941 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3001 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3061 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3661 gtt
//
