LOCUS       sequence-446900-        4008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A Gateway compatible 3' entry clone containing an H2A mCherry fusion
            followed by SV40 late polyA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4008)
  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 4008)
  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: p3E-H2A_mCherry_SV40pA (JDW 967)
FEATURES             Location/Qualifiers
     source          1..4008
                     /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    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1147..1857
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1292..1310)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1560..1579
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1754..1777
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1920..2054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1944..1963)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1998..2017
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2117..2212)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2250..2268)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2273..2289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2273..2289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2391..2410)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2402..3211
                     /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(2469..2488)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3358..3946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3847..3866
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtgggat cctaggatgg caggtggaaa agcaggtaaa
      781 gacagtggaa aagccaagac gaaagccgtc tccagatctc agagagctgg tctgcagttc
      841 cctgtaggtc gtatccacag gcacctgaag acacgaacca ccagccatgg ccgtgtcgga
      901 gctacagctg cggtttacag tgctgctatc ctggagtatc tcaccgctga ggtgctggag
      961 ttggctggaa acgcttctaa ggatctgaag gtgaagcgaa tcactcccag acatttacag
     1021 ctggccatcc gcggagacga ggagctggac tcgcttatca aggccaccat tgctggagga
     1081 ggagtcattc ctcacatcca caagtctctg attggtaaga agggccagca gaaaaccgca
     1141 ggggggatgg tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc
     1201 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1261 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1321 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1381 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1441 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1501 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1561 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1621 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1681 gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1741 aacgtcaaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1801 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaatga
     1861 tagctcgagc tcaagcttcg aattctgcag tctagaacta tagtgagtcg tattacgtag
     1921 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1981 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2041 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2101 tgtgggaggt tttttacaac tttattatac aaagttggca ttataaaaaa gcattgctta
     2161 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggagctcc
     2221 atggtagcgt taacgcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
     2281 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2341 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2401 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2461 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2521 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2581 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2641 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2701 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2761 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2821 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2881 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2941 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3001 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3061 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3121 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3181 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3241 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3301 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3481 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
