LOCUS       sequence-220619-        3268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway entry clone for adding N-terminal fusions of 
            mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3268)
  AUTHORS   Hall TE, Martel N, Lo HP, Xiong Z, Parton RG
  TITLE     A plasmid library of full-length zebrafish rab proteins for in vivo 
            cell biology.
  JOURNAL   Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 
            10.1080/21592799.2017.1301151. eCollection 2017.
  PUBMED    28396820
REFERENCE   2  (bases 1 to 3268)
  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: "Cell 
            Logist."; date: "2017-03-1"; volume: "7(1):e1301151. doi"; pages: " 
            10.1080/21592799.2017.1301151. eCollection 2017"
COMMENT     Sequence Label: pME-mVenus-NS
FEATURES             Location/Qualifiers
     source          1..3268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..44
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     36..53
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     37..53
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    69..168
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      169..178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(175..177,178..180,181..891)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(220..241)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(481..500)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     828..849
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(893..992)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1009..1028)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1010..1028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1033..1049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1033..1049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1151..1170)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1162..1971
                     /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(1229..1248)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2118..2706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2607..2626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2860..2877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3036..3063)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3155..3241)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc
       61 tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
      121 atgagcaatg cttttttata atgccaactt tgtacaaaaa agcaggctgc caccatggtg
      181 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      241 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      301 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      361 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      421 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      481 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      541 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      601 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
      661 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
      721 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      781 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      841 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gcacccagct
      901 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
      961 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1021 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1081 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1141 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1201 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1261 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1321 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1381 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1441 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1501 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1561 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1621 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1681 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1741 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1801 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1861 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1921 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     1981 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2041 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2821 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2881 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2941 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3001 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3061 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3121 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3181 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3241 gatgcctggc agttccctac tctcgcgt
//
