LOCUS       sequence-400936-        8039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ecdysone-inducible vector with SB ITRs for expressing DBEcR (3xmyc- 
            tagged) and FLAG- EGFP; blasticidin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8039)
  AUTHORS   Yeung TK, Kim S, Ma HT, Poon RYC
  TITLE     A robust dual gene ON-OFF toggle directed by two independent 
            promoter-degron pairs.
  JOURNAL   J Cell Sci. 2023 Apr 15;136(8):jcs260754. doi: 10.1242/jcs.260754. 
            Epub 2023 Apr 19.
  PUBMED    36995025
REFERENCE   2  (bases 1 to 8039)
  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: "J Cell 
            Sci."; date: "2023-04-15"; volume: "136(8):jcs260754. doi"; pages: "
            10.1242/jcs.260754. Epub 2023 Apr 19"
COMMENT     Sequence Label: EGFP in pDBEcR-pIND(SP1)/Bla
FEATURES             Location/Qualifiers
     source          1..8039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    313..537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     526..543
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(595..993)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(997..1050)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(1081..1110)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1117..1146)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1153..1182)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2563..2796)
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     regulatory      2796..2805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2831..4013)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(2863..2883)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        4327..4565
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     4512..4533
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             4621..4644
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4651..4674
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4675..4677,4678..4680,4681..5394)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4675..4677,4678..4680,4681..5391)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4720..4741)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4981..5000)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5328..5349
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5513..5532)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5578..5633
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5632..5651)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6025..6042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6196..6784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6276..6295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6955..7746,7747..7815))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7578..7597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7816..7920)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagttgaagt cggaagttta catacactta agttggagtc attaaaactc gtttttcaac
       61 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
      121 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
      181 attcactgta tcacaattcc agtgggtcag aagtttacat acactaagtt cgactcctct
      241 gcagaatgcg gcgatgtttc ggtaaggggt ccgctatcta gtaggcccag ctggttcttt
      301 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
      361 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
      421 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      481 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      541 aggctgatca gcgagctcta gagaattgat ccccaagctt cggccacgaa gtgcttagcc
      601 ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg gctgtccatc
      661 actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct gtcggcaccg
      721 tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac aagtcaggtt
      781 gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag gtcccccagt
      841 aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag ctgcgctggc
      901 gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc tttcaatgag
      961 ggtggattct tcttgagaca aaggcttggc catcgatggg ccaggattct cctcgacgtc
     1021 accgcatgtt agcagacttc ctctgccctc tccactacca gacttgctag tggatccgtt
     1081 caagtcttct tctgagatta atttttgttc accgttcaag tcttcctcgg agattagctt
     1141 ttgttcaccg ttcaaatctt cttcagaaat caacttttgt tcaccacttg cggcagaacc
     1201 ggatcccagc accacggggt tggtgggggg cagcacggtg gggtgggtgg tggccacctc
     1261 ggccacgtcc cagatctcct ccaggaaggg gggcagcttg cggttcttca gcttcaggct
     1321 gatgcacatg ttgctgttct gggtgcccag ggtgcgcagc tcggtcagca cgctcaggat
     1381 gcggccgtag atcacggcgc agcggctgct ggcgctgttc tggttgatga tgtagatgcg
     1441 cagggtgttc aggtagtagc gctggatctc ctccaccagg ctgggctgct ccaggccggg
     1501 gcggtcgctg aagatcacga tggcggtcag cagggcgaag tgcacgttgt ccatgcccat
     1561 cgcgaacatg cagcggcaga agtgcagcag gtcctcgatc acgtaggcca tgccgccctg
     1621 gcggtagttg tcgcgggtgt aggccttgtt gttggcgaac agcacgctgt cgctggcggc
     1681 gtcgtagcgg cgggccacgc gcagcatcat cacctcgctg ctgctggcct tcagcagggt
     1741 gatctggtcg ctctggctga tcttgctgaa gccgggcagg cccttggcga actccacgat
     1801 cagctgcacg gtcaggatgg tcatctcggt gatctggcgg aagggcaggt cgctctcctc
     1861 gtcctcctcg tcgctctgcc aggtctgggt cacgcgcttc aggtcctcgt cgctgggctg
     1921 ctcgtagccc tcctggtacc acaccaggcg ggcgatcagg ctcttctggt tggcgctcag
     1981 gggggggatg ttcttctggc ggttctgctc catcagcttc tcgctcaggt agcggggcac
     2041 cacctcgtgg atgcgggcgg cctcnggggg gggggggtcg cactgcatga tggggggcat
     2101 gtggtcctcc acggtggtgg tgctcacggg cagcaggatg cccttgtcct tcttctggcg
     2161 ctggcggtcc ttgttcttgc tgggctcctg tatgacgcat tccggccgca tacccacggc
     2221 caggcacttt ttcaggcggc actcctgaca ctttcgcctc atgtacatgt ccatttcgca
     2281 ggcgcgcccg aacttgcagc agtagacggc gctcttcgta acgctgcgtc gaaagaacac
     2341 cttgcaggat ccacaggtga gggcgttgta gtggtagccg gaggccctgt cgccgcaaac
     2401 caggcacagc tcctcttgca cccgtggcgc aggtcccttc ttgctcttct tcgcatcgca
     2461 gctttcgttc gccgagtatc cgttcaagct gctcgaaggc gagagatcat cgcgacctga
     2521 agatatagaa tttgatattc ttctagaggt acctagaagc ttcccaccgt actcgtcaat
     2581 tccaagggca tcggtaaaca tctgctcaaa ctcgaagtcg gccatatcca gagcgccgta
     2641 gggggcggag tcgtgggggg taaatcccgg acctggggaa tccccgtccc ccaacatgtc
     2701 cagatcgaaa tcgtctagcg cgtcggcatg cgccatcgcc acgtcctcgc cgtctaagtg
     2761 gagttcgtcc cccaggctga catcggtcgg gggggccatg gtggcctcag aggccagctt
     2821 ggggtagttt tcacgacacc ttaaatggaa gaaaaaaact ttgaaccact gtctgaggct
     2881 tgagaatgaa ccaagatcca aactcaaaaa gggcaaattc caaggagaat tacatcaagt
     2941 gccaagctgg cctaacttca gtctccaccc actcagtatg gggaaactcc atcgcataaa
     3001 acccctcccc ccaacctaaa gacgacgtac tccaaaagct cgagaactaa tcgaggtgcc
     3061 tggacggcgc ccggtactcc gtggagtcac atgaagcgac ggctgaggac ggaaaggccc
     3121 ttttcctttg tgtgggtgac tcacccgccc gctctcccga gcgccgcgtc ctccattttg
     3181 agctccctgc agcagggccg ggaagcggcc atctttccgc tcacgcaact ggtgccgacc
     3241 gggccagcct tgccgcccag ggcggggcga tacacggcgg cgcgaggcca ggcaccagag
     3301 caggccggcc agcttgagac tacccccgtc cgattctcgg tggccgcgct cgcaggcccc
     3361 gcctcgccga acatgtgcgc tgggacgcac gggccccgtc gccgcccgcg gccccaaaaa
     3421 ccgaaatacc agtgtgcaga tcttggcccg catttacaag actatcttgc cagaaaaaaa
     3481 gcgtcgcagc aggtcatcaa aaattttaaa tggctagaga cttatcgaaa gcagcgagac
     3541 aggcgcgaag gtgccaccag attcgcacgc ggcggcccca gcgcccaagc caggcctcaa
     3601 ctcaagcacg aggcgaaggg gctccttaag cgcaaggcct cgaactctcc cacccacttc
     3661 caacccgaag ctcgggatca agaatcacgt actgcagcca ggggcgtgga agtaattcaa
     3721 ggcacgcaag ggccataacc cgtaaagagg ccaggcccgc gggaaccaca cacggcactt
     3781 acctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt
     3841 atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc
     3901 cagtttaccc cgcgccacct tctctaggca ccggttcaat tgccgacccc tccccccaac
     3961 ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gcctctaggg
     4021 tcgacctcga cggatcggga gatctcggcc gcatattaag tgcattgttc tcgataccgc
     4081 taagtgcatt gttctcgtta gctcgatgga caagtgcatt gttctcttgc tgaaagctcg
     4141 atggacaagt gcattgttct cttgctgaaa gctcgatgga caagtgcatt gttctcttgc
     4201 tgaaagctcg atggacaagt gcattgttct cttgctgaaa gctcagtacc cgggtcggag
     4261 tactgccccg cccctagcga ttagccccgg ccccgcatag ctccgccccg ggagtaccct
     4321 cgaccgccgg agtataaata gaggcgcttc gtctacggag cgacaattca attcaaacaa
     4381 gcaaagtgaa cacgtcgcta agcgaaagct aagcaaataa acaagcgcag ctgaacaagc
     4441 taaacaatct gcagtaaagt gcaagttaaa gtgaatcaat taaaagtaac cagcaaccaa
     4501 gtaaatcaac tgcaactact gaaatctgcc aagaagtaat tattgaatac aagaagagaa
     4561 ctctgaatac tttcaacaag ttaccgagaa agaagaactc acacacagct agccacaatg
     4621 gactacaaag acgatgacga taaagccatg ctggaagttc tgtttcaggg gcccatggtg
     4681 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4741 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     4801 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4861 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     4921 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     4981 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     5041 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     5101 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     5161 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     5221 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     5281 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5341 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     5401 cgcgactcta gaggatccag acatgataag aattcactcc tcaggtgcag gctgcctatc
     5461 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     5521 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     5581 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     5641 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     5701 caacatatgc catatgctgg ctgccatgaa ctagctactc gggacccctt accgaaacat
     5761 cgccgcattc tgcagaggag tcgagtgtat gtaaacttct gacccactgg gaatgtgatg
     5821 aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat ttcacattct
     5881 taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag gattaaatgt
     5941 caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta aacttccgac
     6001 ttcaactgta tagggatccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7921 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
     7981 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattgg atccctata
//
