LOCUS       sequence-338403-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of TIR1-myc and tTA in sleeping beauty 
            transposon vector with selection marker (zeocin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  AUTHORS   Yeung TK, Lau HW, Ma HT, Poon RYC
  TITLE     One-step multiplex toolkit for efficient generation of conditional 
            gene silencing human cell lines.
  JOURNAL   Mol Biol Cell. 2021 May 12:mbcE21020051. doi: 
            10.1091/mbc.E21-02-0051.
  PUBMED    33979199
REFERENCE   2  (bases 1 to 8098)
  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: "Mol Biol 
            Cell. 2021 May 12:mbcE21020051. doi: 10.1091/mbc.E21-02-0051."
COMMENT     Sequence Label: pSBbi-TIR1-tTA/Zeo
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(383..400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(554..1142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(634..653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1313..2104,2105..2173))
                     /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     1936..1955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2174..2278)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2710..2934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2923..2940
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2997..3026)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3033..3062)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3069..3098)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3126..4850)
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MAYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     regulatory      4847..4856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4882..6064)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(4914..4934)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(6765..6784)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             7319..7429
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7442..7495
                     /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             7499..7873
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVSDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(7969..7988)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8034..8089
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(join(8088..8098,1..9))
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
       61 acatatgcca tatgctggct gccatgaact agctactcgg gaccccttac cgaaacatcg
      121 ccgcattctg cagaggagtc gagtgtatgt aaacttctga cccactggga atgtgatgaa
      181 agaaataaaa gctgaaatga atcattctct ctactattat tctgatattt cacattctta
      241 aaataaagtg gtgatcctaa ctgacctaag acagggaatt tttactagga ttaaatgtca
      301 ggaattgtga aaaagtgagt ttaaatgtat ttggctaagg tgtatgtaaa cttccgactt
      361 caactgtata gggatccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      421 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      481 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      541 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      601 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      661 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      721 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      781 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      841 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      901 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      961 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1021 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1081 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1261 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1321 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1381 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1441 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1501 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1561 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1621 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1681 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1741 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1801 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1861 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1921 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1981 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2041 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2101 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2161 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2221 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2281 catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2341 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattggat ccctatacag
     2401 ttgaagtcgg aagtttacat acacttaagt tggagtcatt aaaactcgtt tttcaactac
     2461 tccacaaatt tcttgttaac aaacaatagt tttggcaagt cagttaggac atctactttg
     2521 tgcatgacac aagtcatttt tccaacaatt gtttacagac agattatttc acttataatt
     2581 cactgtatca caattccagt gggtcagaag tttacataca ctaagttcga ctcctctgca
     2641 gaatgcggcg atgtttcggt aaggggtccg ctatctagta ggcccagctg gttctttccg
     2701 cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct
     2761 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     2821 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
     2881 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg
     2941 ctgatcagcg agctctagag aattgatccc caagcttttt agctagtgga tccgttcaag
     3001 tcttcttctg agattaattt ttgttcaccg ttcaagtctt cctcggagat tagcttttgt
     3061 tcaccgttca aatcttcttc agaaatcaac ttttgttcac cactagcagc agaaccggag
     3121 tcgactagga ttttaacaaa atttggtgca tcatccctcg ccccagcagt tgtgcggtac
     3181 acatataatt tctccacctt tggcaggtcg ccatgatttt cctccatttc gttagacccg
     3241 tcccgctcat taatgacttc cacgttcagc ataggcattt tactagccag gacctgacag
     3301 cccttcagag tcacattgca actagacatc cacaggctcc tcattgtctc gtatctggca
     3361 aagtttccca gcagagcggc atccccgaaa gggctgtccc ggatttccag ttttcgcaga
     3421 ttcttacacc cgttcatcac gtgcatcatg cccttatcgg agtctccggc aaatgcaatg
     3481 ctcagcatct ccagctgctt agcatacttg ccgatgtaca tgaacacttt gtctgtcagc
     3541 aggccggaaa tgctcagcct ctggagtccc ttgcactctc tcacgatagc tccaaaccct
     3601 tcatccagtg gctgggatgt gaccacgtcg ggtttgcctg gttccagaat acacagccga
     3661 aatcgggtga aattagggca gttcttggcg acggtcacca gtgcagcatt agtcatctga
     3721 tgacagaaat acagcaggct gttcagcttt gggcatccca gtgagactgc caccaggcct
     3781 tcctcagtga cagcactata tccggccacg taaaaatcag atgggaacac tctcagctcc
     3841 tggaggtcct tgcaggagct ggcgaccacc tggagcccct tatcggaaat acagtccagc
     3901 acccacagtc gctggagttt cacgcagcgt gagatcatct ttgtcagatc actggcgtcc
     3961 agggttggtg catagctcag gttcaggcct gtcagctgtg cacacagagg gtagataaag
     4021 ctcaggcaca cgggtgaagc atcccagaat cctgacagac tcctcagcat tttacacttc
     4081 tccagggcag aagtcagttt aaagtaggat tctgtctgga aatcgtctgt cagattgccg
     4141 gttcccagat cctccaggtt aggggttctc agcagaatct tagccagagt gtccacgctc
     4201 acgctccggt tcagccgcag agacctcagg ttggggcttc ttgagaccag ccgctccagg
     4261 gacccagcat tcacttcgcc tttgatgcag gcaaagttca gggacaccag gctggtacat
     4321 gagtctggga agcaagacag ccatctgggg cctctatcct ccacttcatt ctcctggagg
     4381 tccagctccc gcagcagctt acagtgggag gccacggctg ccagtccatc ggtgctgaag
     4441 ccttcgcaag aaatcagcac cagtgcgcga aacctaggga agctccgagc cagcagctcc
     4501 agactttcgt cagagaccac cattcgcttc atgcgcagtt cctccagtcc atgacagccg
     4561 cgagcggctg cctcgatcca agggccggcg tatcctcccc aatcaggggg caccaggtta
     4621 aagtcggcga agtggggttt ccccttgacg gtcagtgcgc gcacatttgg aaacctagcg
     4681 gcgaccctgc ctgctctcac agcatagcag ttgccgacga acactcctct ccggctcagg
     4741 cgttcgatct cgtaccacac tttgcagacc aggctcactg tgtttctatc tctctgtgca
     4801 ggcaggaagc taaaaatgtg ttcgacgacc tcttcaggaa agtatgccat ggtggcctca
     4861 gaggccagct tggggtagtt ttcacgacac cttaaatgga agaaaaaaac tttgaaccac
     4921 tgtctgaggc ttgagaatga accaagatcc aaactcaaaa agggcaaatt ccaaggagaa
     4981 ttacatcaag tgccaagctg gcctaacttc agtctccacc cactcagtat ggggaaactc
     5041 catcgcataa aacccctccc cccaacctaa agacgacgta ctccaaaagc tcgagaacta
     5101 atcgaggtgc ctggacggcg cccggtactc cgtggagtca catgaagcga cggctgagga
     5161 cggaaaggcc cttttccttt gtgtgggtga ctcacccgcc cgctctcccg agcgccgcgt
     5221 cctccatttt gagctccctg cagcagggcc gggaagcggc catctttccg ctcacgcaac
     5281 tggtgccgac cgggccagcc ttgccgccca gggcggggcg atacacggcg gcgcgaggcc
     5341 aggcaccaga gcaggccggc cagcttgaga ctacccccgt ccgattctcg gtggccgcgc
     5401 tcgcaggccc cgcctcgccg aacatgtgcg ctgggacgca cgggccccgt cgccgcccgc
     5461 ggccccaaaa accgaaatac cagtgtgcag atcttggccc gcatttacaa gactatcttg
     5521 ccagaaaaaa agcgtcgcag caggtcatca aaaattttaa atggctagag acttatcgaa
     5581 agcagcgaga caggcgcgaa ggtgccacca gattcgcacg cggcggcccc agcgcccaag
     5641 ccaggcctca actcaagcac gaggcgaagg ggctccttaa gcgcaaggcc tcgaactctc
     5701 ccacccactt ccaacccgaa gctcgggatc aagaatcacg tactgcagcc aggggcgtgg
     5761 aagtaattca aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac
     5821 acacggcact tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga
     5881 ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga
     5941 catcactttc ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc
     6001 ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg
     6061 agcctctagg cgagaccctg tctcacaaaa taaagtaagc ccggactgag tgcggaaagg
     6121 cgggcctggc gggtctggtc tccccatgcg ggccaccaga ggccctgcag ccttcagtcg
     6181 cttgaagggg taatggcgct tccactcaca aacatggcgg acagagcgtg tgaacgagat
     6241 gaacagcccc tcaaaaatat ggccgccgag gctggacggc cgtgccccag cagcaccgcc
     6301 tccgcgcccc acgtgatctc tcgccgggca cagcgctgac cgcggaggtc caaccggaag
     6361 aatgtccgga ttggacattc ggaagagggc ccgccttccc tggggaatct ctgcgcacgc
     6421 gcagaacgct tcgaccaatg aaaacacagg aagccgtccg cgcaaccgcg ttgcgtcact
     6481 tctgccgccc ctgtttcaag gtatatagcc gtagacggaa cttcgccttt ctctcggcct
     6541 tagcgccatt tttttgggtg agtgtttttt ggttcctgcg ttgggattcc gtgtacaatc
     6601 catagacatc tgacctcggc acttagcatc atcacagcaa actaactgta gcctttctct
     6661 ctttccctgt agaaacctct gcactcgaca tatgtctaga ttagataaaa gtaaagtgat
     6721 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact
     6781 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc
     6841 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt
     6901 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt
     6961 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta
     7021 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc
     7081 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga
     7141 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt
     7201 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct
     7261 tgaattgatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga
     7321 cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg actttgacct
     7381 tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccggatctgg
     7441 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccatcgat
     7501 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     7561 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     7621 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgtcggacaa
     7681 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
     7741 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
     7801 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     7861 ggagcaggac tgacacgtgc tacgagaatt cactcctcag gtgcaggctg cctatcagaa
     7921 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     7981 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     8041 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaagg
//
