LOCUS       sequence-484484-        4651 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Gateway middle entry clone containing EGFP fused to oncogenic c-Myc.
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4651)
  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 4651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 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-EGFP-Myc (JDW 1441)
FEATURES             Location/Qualifiers
     source          1..4651
                     /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             join(704..706,707..709,710..1420)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(704..706,707..709,710..1420)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(749..770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1010..1029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1357..1378
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1436..2755
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     protein_bind    complement(2776..2875)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2892..2911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2893..2911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2916..2932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2916..2932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3034..3053)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3045..3854
                     /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(3112..3131)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4001..4589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4490..4509
                     /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 agcaggcttc agtgactagt taggtaccta ttaattaagg ccgatggtga gcaagggcga
      721 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      781 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      841 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      901 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      961 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1021 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1081 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1141 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1201 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1261 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1321 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1381 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtggaa gatctatgcc
     1441 cctcaacgtt agcttcacca acaggaacta tgacctcgac tacgactcgg tgcagccgta
     1501 tttctactgc gacgaggagg agaacttcta ccagcagcag cagcagagcg agctgcagcc
     1561 cccggcgccc agcgaggata tctggaagaa attcgagctg ctgcccaccc cgcccctgtc
     1621 ccctagccgc cgctccgggc tctgctcgcc ctcctacgtt gcggtcacac ccttctccct
     1681 tcggggagac aacgacggcg gtggcgggag cttctccacg gccgaccagc tggagatggt
     1741 gaccgagctg ctgggaggag acatggtgaa ccagagtttc atctgcgacc cggacgacga
     1801 gaccttcatc aaaaacatca tcatccagga ctgtatgtgg agcggcttct cggccgccgc
     1861 caagctcgtc tcagagaagc tggcctccta ccaggctgcg cgcaaagaca gcggcagccc
     1921 gaaccccgcc cgcggccaca gcgtctgctc cacctccagc ttgtacctgc aggatctgag
     1981 cgccgccgcc tcagagtgca tcgacccctc ggtggtcttc ccctaccctc tcaacgacag
     2041 cagctcgccc aagtcctgcg cctcgcaaga ctccagcgcc ttctctccgt cctcggattc
     2101 tctgctctcc tcgacggagt cctccccgca gggcagcccc gagcccctgg tgctccatga
     2161 ggagacaccg cccaccacca gcagcgactc tgaggaggaa caagaagatg aggaagaaat
     2221 cgatgttgtt tctgtggaaa agaggcaggc tcctggcaaa aggtcagagt ctggatcacc
     2281 ttctgctgga ggccacagca aacctcctca cagcccactg gtcctcaaga ggtgccacgt
     2341 ctccacacat cagcacaact acgcagcgcc tccctccact cggaaggact atcctgctgc
     2401 caagagggtc aagttggaca gtgtcagagt cctgagacag atcagcaaca accgaaaatg
     2461 caccagcccc aggtcctcgg acaccgagga gaatgtcaag aggcgaacac acaacgtctt
     2521 ggagcgccag aggaggaacg agctaaaacg gagctttttt gccctgcgtg accagatccc
     2581 ggagttggaa aacaatgaaa aggcccccaa ggtagttatc cttaaaaaag ccacagcata
     2641 catcctgtcc gtccaagcag aggagcaaaa gctcatttct gaagaggact tgttgcggaa
     2701 acgacgagaa cagttgaaac acaaacttga acagctacgg aactcttgtg cgtaataact
     2761 agcggccgca gtcagaccca gctttcttgt acaaagttgg cattataaga aagcattgct
     2821 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     2881 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2941 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     3001 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     3061 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     3121 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     3181 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     3241 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     3301 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     3361 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     3421 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     3481 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3541 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3601 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3661 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3721 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3781 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3841 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     3901 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4141 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4381 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4621 tggccttttg ctggcctttt gctcacatgt t
//
