LOCUS       sequence-274737-        4497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides IRES-Venus-pA cassette on Gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 4497)
  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-R2L3-IRES-Venus-pA
FEATURES             Location/Qualifiers
     source          1..4497
                     /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"
     misc_feature    join(763..1337,1338..1340,1341..1349)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(930..947)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1157..1176
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1350..1352,1353..1355,1356..2069)
                     /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(1395..1416)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1656..1675)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2003..2024
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2159..2178)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2224..2279
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2278..2297)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(2606..2701)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2738..2757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2739..2757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2762..2778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2762..2778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2880..2899)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2891..3700
                     /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(2958..2977)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3847..4435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4336..4355
                     /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 aaagtgggaa ttccgccccc cccccctctc cctccccccc
      781 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
      841 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
      901 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
      961 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     1021 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     1081 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     1141 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     1201 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     1261 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     1321 aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg
     1381 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1441 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
     1501 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt
     1561 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1621 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1681 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1741 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1801 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1861 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1921 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     1981 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2041 ctctcggcat ggacgagctg tacaagtaaa gcggccaatt cactcctcag gtgcaggctg
     2101 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     2161 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2221 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2281 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     2341 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     2401 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     2461 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     2521 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     2581 ctgtccctct tctcttatgg agatccaact ttattataca aagttggcat tataaaaaag
     2641 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     2701 tggagctcca tggtagcgtt aacgcggccg cgatatcccc tatagtgagt cgtattacat
     2761 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2821 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2881 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2941 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3001 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3061 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3121 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3181 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3241 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3301 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3361 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3421 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3481 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3541 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3601 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3661 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3721 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3781 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3841 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3901 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3961 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4021 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4081 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4141 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4201 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4261 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4321 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4381 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4441 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
