LOCUS       sequence-274619-        3269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Venus ORF with stop codon between attL1-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3269)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3269)
  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"
COMMENT     Sequence Label: pENTR-Venus
FEATURES             Location/Qualifiers
     source          1..3269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(23..25,26..28,29..742)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(68..89)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(329..348)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     676..697
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(743..842)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(859..878)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(860..878)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(883..899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(883..899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1001..1020)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1012..1821
                     /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(1079..1098)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1968..2556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2457..2476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2710..2727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2886..2913)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3005..3091)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3140..3162
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3154..3171
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3155..3171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3187..3269,1..17)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtacaaaaaa gcaggctcca ccatggtgag caagggcgag gagctgttca ccggggtggt
       61 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      121 gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa
      181 gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc
      241 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      301 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      361 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      421 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      481 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
      541 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      601 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
      661 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      721 catggacgag ctgtacaagt aaacccagct ttcttgtaca aagttggcat tataagaaag
      781 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      841 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
      901 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
      961 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1021 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1081 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1141 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1201 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1261 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1321 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1381 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1441 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1501 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1561 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     1621 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     1681 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     1741 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     1801 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     1861 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     1921 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1981 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2041 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2101 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2161 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2221 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2281 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2341 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2401 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2461 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2521 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2581 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2641 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2701 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     2761 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     2821 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     2881 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     2941 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     3001 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     3061 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     3121 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     3181 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
     3241 gagcaatgct tttttataat gccaacttt
//
