LOCUS       sequence-338226-        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-controlled AID-fusion sleeping beauty transposon vector with 
            selection marker (blasticidin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  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 5387)
  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: pUHD-SB-AID/Bla
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(311..329,330..352,353..371,372..394,395..413,414..436,
                     437..455,456..478,479..497,498..520,521..539,540..562,
                     563..581)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        614..652
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     649..673
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             748..1431
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /label=auxin-inducible degron
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="MGSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNN
                     EPANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSG
                     GPEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMN
                     ERKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAME
                     KCKSRA"
     polyA_signal    1545..1666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1556..1575)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1610..1629
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2383..2781
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(2833..2850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2839..3063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3389..3406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3560..4148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3640..3659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4319..5110,5111..5179))
                     /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     4942..4961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5180..5284)
                     /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 ggccctttcg tctttagtcg
      301 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
      361 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
      421 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
      481 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
      541 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg
      601 gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt
      661 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagagaccg
      721 ggaccgatcc agcctccgcg ggctagcatg ggcagtgtcg agctgaatct gagggagact
      781 gagctgtgtc ttggtcttcc cggtggagat acagtggctc cggtaaccgg aaacaagaga
      841 gggttctcag agacggttga tctgaagcta aatctgaata atgagcctgc aaacaaggaa
      901 ggatctacga ctcatgacgt cgtgactttt gattccaagg agaagagtgc ttgtcctaaa
      961 gatccagcca aacctccggc caaggcacaa gttgtgggat ggccaccggt gagatcatac
     1021 cggaagaacg tgatggtttc ctgccaaaaa tcaagcggtg gcccggaggc ggcggcgttc
     1081 gtgaaggtat caatggacgg agcaccgtac ttgaggaaaa tcgatttgag gatgtataaa
     1141 agctacgatg agctttctaa tgctttgtcc aacatgttca gctcttttac gatgggcaaa
     1201 catggaggag aagaaggaat gatagacttc atgaatgaga ggaaattgat ggatttggtg
     1261 aatagctggg actatgttcc ctcttatgaa gacaaagacg gtgattggat gctcgtcggc
     1321 gacgttcctt ggccaatgtt cgtcgataca tgcaagcgtt tacgtctcat gaaaggatcg
     1381 gatgccattg gtctcgctcc gagggcgatg gagaagtgca agagcagagc tgcggccgga
     1441 gctggtgcag gcgctggagc gggtgccgat atcctattca ccatggctcg agaattcgag
     1501 ctcggtaccc ggggatccag acatgataag atcgatagac atgataagat acattgatga
     1561 gtttggacaa accacaacaa gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1621 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1681 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     1741 cctctacaaa tgtggtacct agacgagacc ctgtctcaca aaataaagta agcccggact
     1801 gagtgcggaa aggcgggcct ggcgggtctg gtctccccat gcgggccacc agaggccctg
     1861 cagccttcag tcgcttgaag gggtaatggc gcttccactc acaaacatgg cggacagagc
     1921 gtgtgaacga gatgaacagc ccctcaaaaa tatggccgcc gaggctggac ggccgtgccc
     1981 cagcagcacc gcctccgcgc cccacgtgat ctctcgccgg gcacagcgct gaccgcggag
     2041 gtccaaccgg aagaatgtcc ggattggaca ttcggaagag ggcccgcctt ccctggggaa
     2101 tctctgcgca cgcgcagaac gcttcgacca atgaaaacac aggaagccgt ccgcgcaacc
     2161 gcgttgcgtc acttctgccg cccctgtttc aaggtatata gccgtagacg gaacttcgcc
     2221 tttctctcgg ccttagcgcc atttttttgg gtgagtgttt tttggttcct gcgttgggat
     2281 tccgtgtaca atccatagac atctgacctc ggcacttagc atcatcacag caaactaact
     2341 gtagcctttc tctctttccc tgtagaaacc tctgcactcg acatggccaa gcctttgtct
     2401 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     2461 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     2521 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     2581 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     2641 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     2701 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     2761 ggttatgtgt gggagggcta agcacttcgt ggccgaagct tggggatcaa ttctctagag
     2821 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2881 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2941 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3001 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3061 tggcttctga ggcggaaaga accagctggg cctactagct actcgggacc ccttaccgaa
     3121 acatcgccgc attctgcaga ggagtcgagt gtatgtaaac ttctgaccca ctgggaatgt
     3181 gatgaaagaa ataaaagctg aaatgaatca ttctctctac tattattctg atatttcaca
     3241 ttcttaaaat aaagtggtga tcctaactga cctaagacag ggaattttta ctaggattaa
     3301 atgtcaggaa ttgtgaaaaa gtgagtttaa atgtatttgg ctaaggtgta tgtaaacttc
     3361 cgacttcaac tgtataggga tctgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3421 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3481 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3541 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3601 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3661 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3721 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3781 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3841 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3901 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3961 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4021 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4081 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4141 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4201 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4261 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4321 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4381 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4441 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4501 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4561 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4621 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4681 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4741 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4801 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4861 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4921 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4981 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5041 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5101 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5161 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5221 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5281 cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta
     5341 aggagaaaat accgcatcag gaaattgtaa gcgttaagat ccctata
//
