LOCUS       sequence-447428-        4882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone containing the CAG promoter for 
            constitutive expression of downstream constructs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4882)
  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 4882)
  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: p5E-CAGGS (JDW 912)
FEATURES             Location/Qualifiers
     source          1..4882
                     /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    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        690..708
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     734..750
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        898..1277
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2669..2691
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2753..2772
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2856..2872)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2856..2872)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2909..2927)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2928..3051
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3124..3142)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3147..3163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3147..3163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3265..3284)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3276..4085
                     /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(3343..3362)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4232..4820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4721..4740
                     /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 tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggt aatacgactc actatagggc gaattgggta
      721 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagccca
      781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac
      901 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      961 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1021 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1081 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1141 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     1201 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     1261 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     1321 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     1381 gggcgggggg gggggggggg ggggcgccgg ggggggcggg gggggggggg gggggggggg
     1441 gggggggcgg gggggggggg gggggggggg gggggggggg gggggggggg ggggggcggt
     1501 gatgtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     1561 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     1621 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     1681 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     1741 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
     1801 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     1861 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
     1921 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     1981 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     2041 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     2101 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     2161 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggggg ccccggggcg
     2221 gggggggggg gggggggggg gggggggcgg cgcgcggggg gggggggggg gggggggggg
     2281 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gcggccgcgg
     2341 gggggggggg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     2401 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     2461 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     2521 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     2581 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     2641 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     2701 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     2761 ctggttattg tgctgtctca tcattttggc aaagaattgc tcgaggtcga tcgacggtat
     2821 cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc
     2881 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattcaa gtttgtacaa
     2941 aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     3001 taaaaaacag actacataat actgtaaaac acaacatatg cagtcactat gaaccaacta
     3061 cttagatggt attagtgacc tgtagaattc gagctctaga gctgcagggc ggccgcgata
     3121 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     3181 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3241 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3301 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3361 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     3421 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3481 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3541 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     3601 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3661 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     3721 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     3781 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     3841 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     3901 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     3961 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     4021 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     4081 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     4141 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     4201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4861 gctggccttt tgctcacatg tt
//
