LOCUS       sequence-274673-        3395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  d4Venus 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 3395)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3395)
  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-d4Venus
FEATURES             Location/Qualifiers
     source          1..3395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(23..25,26..28,29..739)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVCKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     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(869..968)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(985..1004)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(986..1004)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1009..1025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1009..1025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1127..1146)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1138..1947
                     /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(1205..1224)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2094..2682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2583..2602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2836..2853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3012..3039)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3131..3217)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3266..3288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3280..3297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3281..3297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3313..3395,1..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 ccatggtgtg 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 ctgtacaaga agcttagcca tggcttcccg ccggaggtgg aggagcagga
      781 tgatggcgcg ctgcccatgt cttgtgccca ggagagcggg atggaccgtc accctgcagc
      841 ctgtgcttct gctaggatca atgtgtagac ccagctttct tgtacaaagt tggcattata
      901 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
      961 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     1021 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1081 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1141 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     1201 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     1261 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     1321 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     1381 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     1441 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     1501 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     1561 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     1621 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     1681 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     1741 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     1801 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1861 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1921 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     1981 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     2041 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2221 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2761 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2821 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2881 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2941 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     3001 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     3061 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     3121 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     3181 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     3241 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     3301 aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
     3361 attgatgagc aatgcttttt tataatgcca acttt
//
