LOCUS       sequence-400918-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with SB ITRs for expressing DBEcR (3xmyc-tagged) under 
            EF1-alpha promoter; blasticidin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  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 6636)
  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: pDBEcR/Bla
FEATURES             Location/Qualifiers
     source          1..6636
                     /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"
     primer_bind     complement(4110..4129)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4175..4230
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4229..4248)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4622..4639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4793..5381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4873..4892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5552..6343,6344..6412))
                     /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     6175..6194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6413..6517)
                     /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 cctcgggggg 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 tcgaccgaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     4081 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     4141 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     4201 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     4261 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccat atgctggctg
     4321 ccatgaacta gctactcggg accccttacc gaaacatcgc cgcattctgc agaggagtcg
     4381 agtgtatgta aacttctgac ccactgggaa tgtgatgaaa gaaataaaag ctgaaatgaa
     4441 tcattctctc tactattatt ctgatatttc acattcttaa aataaagtgg tgatcctaac
     4501 tgacctaaga cagggaattt ttactaggat taaatgtcag gaattgtgaa aaagtgagtt
     4561 taaatgtatt tggctaaggt gtatgtaaac ttccgacttc aactgtatag ggatccgctt
     4621 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4681 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4741 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4801 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4861 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4921 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4981 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5041 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5101 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5161 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5221 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5281 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5341 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5401 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5461 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5521 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5581 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5641 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5701 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5761 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5821 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5881 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5941 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6001 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6061 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6121 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6181 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6241 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6301 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6361 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6421 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6481 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6541 ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     6601 taagcgttaa tattttgtta aaattggatc cctata
//
