LOCUS       sequence-338413-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of TIR1-myc in sleeping beauty transposon 
            vector with selection marker (puromycin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7524)
  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 7524)
  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/Pur
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     584..601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     975..994
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    993..1048
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1094..1113
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1213..1812)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1296..1316)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1793..1812
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        2444..3626
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     3574..3594
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3652..3661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3658..5382
                     /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"
     CDS             5410..5439
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5446..5475
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5482..5511
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5568..5585)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5574..5798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6230..6334
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6335..6403,6404..7195)
                     /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     complement(6553..6572)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7366..7524,1..430)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
       61 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      481 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      541 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      601 ggaagcggat ccctatacag ttgaagtcgg aagtttacat acaccttagc caaatacatt
      661 taaactcact ttttcacaat tcctgacatt taatcctagt aaaaattccc tgtcttaggt
      721 cagttaggat caccacttta ttttaagaat gtgaaatatc agaataatag tagagagaat
      781 gattcatttc agcttttatt tctttcatca cattcccagt gggtcagaag tttacataca
      841 ctcgactcct ctgcagaatg cggcgatgtt tcggtaaggg gtcccgagta gctagttcat
      901 ggcagccagc atatggcata tgttgccaaa ctctaaacca aatactcatt ctgatgtttt
      961 aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt ccaacacact
     1021 attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag gggcttcatg
     1081 atgtccccat aatttttggc agagggaaaa agatctcagt ggtatttgtg agccagggca
     1141 ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt gaattcacga
     1201 caggccttcg aatcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
     1261 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
     1321 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
     1381 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
     1441 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
     1501 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
     1561 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
     1621 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
     1681 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
     1741 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
     1801 gtactcggtc atggtggcct caggtgcaga ggtttctaca gggaaagaga gaaaggctac
     1861 agttagtttg ctgtgatgat gctaagtgcc gaggtcagat gtctatggat tgtacacgga
     1921 atcccaacgc aggaaccaaa aaacactcac ccaaaaaaat ggcgctaagg ccgagagaaa
     1981 ggcgaagttc cgtctacggc tatatacctt gaaacagggg cggcagaagt gacgcaacgc
     2041 ggttgcgcgg acggcttcct gtgttttcat tggtcgaagc gttctgcgcg tgcgcagaga
     2101 ttccccaggg aaggcgggcc ctcttccgaa tgtccaatcc ggacattctt ccggttggac
     2161 ctccgcggtc agcgctgtgc ccggcgagag atcacgtggg gcgcggaggc ggtgctgctg
     2221 gggcacggcc gtccagcctc ggcggccata tttttgaggg gctgttcatc tcgttcacac
     2281 gctctgtccg ccatgtttgt gagtggaagc gccattaccc cttcaagcga ctgaaggctg
     2341 cagggcctct ggtggcccgc atggggagac cagacccgcc aggcccgcct ttccgcactc
     2401 agtccgggct tactttattt tgtgagacag ggtctcgcct agaggctccg gtgcccgtca
     2461 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     2521 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     2581 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     2641 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     2701 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cgcccctggc
     2761 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     2821 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggctt gggcgctggg
     2881 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     2941 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3001 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3061 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     3121 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     3181 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     3241 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     3301 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     3361 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     3421 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccata
     3481 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     3541 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     3601 tttttcttcc atttaaggtg tcgtgaaaac taccccaagc tggcctctga ggccaccatg
     3661 gcatactttc ctgaagaggt cgtcgaacac atttttagct tcctgcctgc acagagagat
     3721 agaaacacag tgagcctggt ctgcaaagtg tggtacgaga tcgaacgcct gagccggaga
     3781 ggagtgttcg tcggcaactg ctatgctgtg agagcaggca gggtcgccgc taggtttcca
     3841 aatgtgcgcg cactgaccgt caaggggaaa ccccacttcg ccgactttaa cctggtgccc
     3901 cctgattggg gaggatacgc cggcccttgg atcgaggcag ccgctcgcgg ctgtcatgga
     3961 ctggaggaac tgcgcatgaa gcgaatggtg gtctctgacg aaagtctgga gctgctggct
     4021 cggagcttcc ctaggtttcg cgcactggtg ctgatttctt gcgaaggctt cagcaccgat
     4081 ggactggcag ccgtggcctc ccactgtaag ctgctgcggg agctggacct ccaggagaat
     4141 gaagtggagg atagaggccc cagatggctg tcttgcttcc cagactcatg taccagcctg
     4201 gtgtccctga actttgcctg catcaaaggc gaagtgaatg ctgggtccct ggagcggctg
     4261 gtctcaagaa gccccaacct gaggtctctg cggctgaacc ggagcgtgag cgtggacact
     4321 ctggctaaga ttctgctgag aacccctaac ctggaggatc tgggaaccgg caatctgaca
     4381 gacgatttcc agacagaatc ctactttaaa ctgacttctg ccctggagaa gtgtaaaatg
     4441 ctgaggagtc tgtcaggatt ctgggatgct tcacccgtgt gcctgagctt tatctaccct
     4501 ctgtgtgcac agctgacagg cctgaacctg agctatgcac caaccctgga cgccagtgat
     4561 ctgacaaaga tgatctcacg ctgcgtgaaa ctccagcgac tgtgggtgct ggactgtatt
     4621 tccgataagg ggctccaggt ggtcgccagc tcctgcaagg acctccagga gctgagagtg
     4681 ttcccatctg atttttacgt ggccggatat agtgctgtca ctgaggaagg cctggtggca
     4741 gtctcactgg gatgcccaaa gctgaacagc ctgctgtatt tctgtcatca gatgactaat
     4801 gctgcactgg tgaccgtcgc caagaactgc cctaatttca cccgatttcg gctgtgtatt
     4861 ctggaaccag gcaaacccga cgtggtcaca tcccagccac tggatgaagg gtttggagct
     4921 atcgtgagag agtgcaaggg actccagagg ctgagcattt ccggcctgct gacagacaaa
     4981 gtgttcatgt acatcggcaa gtatgctaag cagctggaga tgctgagcat tgcatttgcc
     5041 ggagactccg ataagggcat gatgcacgtg atgaacgggt gtaagaatct gcgaaaactg
     5101 gaaatccggg acagcccttt cggggatgcc gctctgctgg gaaactttgc cagatacgag
     5161 acaatgagga gcctgtggat gtctagttgc aatgtgactc tgaagggctg tcaggtcctg
     5221 gctagtaaaa tgcctatgct gaacgtggaa gtcattaatg agcgggacgg gtctaacgaa
     5281 atggaggaaa atcatggcga cctgccaaag gtggagaaat tatatgtgta ccgcacaact
     5341 gctggggcga gggatgatgc accaaatttt gttaaaatcc tagtcgactc cggttctgct
     5401 gctagtggtg aacaaaagtt gatttctgaa gaagatttga acggtgaaca aaagctaatc
     5461 tccgaggaag acttgaacgg tgaacaaaaa ttaatctcag aagaagactt gaacggatcc
     5521 actagctaaa aagcttgggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc
     5581 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5641 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5701 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5761 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5821 ctgggcctac tagatagcgg accccttacc gaaacatcgc cgcattctgc agaggagtcg
     5881 aacttagtgt atgtaaactt ctgacccact ggaattgtga tacagtgaat tataagtgaa
     5941 ataatctgtc tgtaaacaat tgttggaaaa atgacttgtg tcatgcacaa agtagatgtc
     6001 ctaactgact tgccaaaact attgtttgtt aacaagaaat ttgtggagta gttgaaaaac
     6061 gagttttaat gactccaact taagtgtatg taaacttccg acttcaactg tatagggatc
     6121 caattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     6181 ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6241 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6301 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6361 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6421 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6481 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6541 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6601 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6661 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6721 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6781 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6841 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6901 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6961 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7021 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7081 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7141 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7201 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7261 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7321 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7381 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7441 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7501 accaaatact gttcttctag tgta
//
