LOCUS       sequence-338406-        4874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-controlled mAID-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 4874)
  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 4874)
  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-mAID/Bla
FEATURES             Location/Qualifiers
     source          1..4874
                     /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             760..963
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     polyA_signal    1032..1153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1043..1062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1097..1116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1870..2268
                     /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(2320..2337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2326..2550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2876..2893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3047..3635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3127..3146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3806..4597,4598..4666))
                     /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     4429..4448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4667..4771)
                     /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 gccggagcaa aggagaaatc agcctgccct
      781 aaagaccccg caaagccccc cgcaaaagca caggtggtgg gatggccccc cgtgcgcagc
      841 taccggaaga acgtgatggt gtcctgccag aagagctccg gaggaccaga ggcagcagcc
      901 tttgtgaagg tgtctatgga tggcgcccct tacctgagaa agatcgacct gaggatgtat
      961 aaggcctcca tggctcgaga attcgagctc ggtacccggg gatccagaca tgataagatc
     1021 gatagacatg ataagataca ttgatgagtt tggacaaacc acaacaagaa tgcagtgaaa
     1081 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1141 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     1201 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacctaga cgagaccctg
     1261 tctcacaaaa taaagtaagc ccggactgag tgcggaaagg cgggcctggc gggtctggtc
     1321 tccccatgcg ggccaccaga ggccctgcag ccttcagtcg cttgaagggg taatggcgct
     1381 tccactcaca aacatggcgg acagagcgtg tgaacgagat gaacagcccc tcaaaaatat
     1441 ggccgccgag gctggacggc cgtgccccag cagcaccgcc tccgcgcccc acgtgatctc
     1501 tcgccgggca cagcgctgac cgcggaggtc caaccggaag aatgtccgga ttggacattc
     1561 ggaagagggc ccgccttccc tggggaatct ctgcgcacgc gcagaacgct tcgaccaatg
     1621 aaaacacagg aagccgtccg cgcaaccgcg ttgcgtcact tctgccgccc ctgtttcaag
     1681 gtatatagcc gtagacggaa cttcgccttt ctctcggcct tagcgccatt tttttgggtg
     1741 agtgtttttt ggttcctgcg ttgggattcc gtgtacaatc catagacatc tgacctcggc
     1801 acttagcatc atcacagcaa actaactgta gcctttctct ctttccctgt agaaacctct
     1861 gcactcgaca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     1921 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     1981 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     2041 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     2101 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     2161 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     2221 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc acttcgtggc
     2281 cgaagcttgg ggatcaattc tctagagctc gctgatcagc ctcgactgtg ccttctagtt
     2341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2521 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctgggcct
     2581 actagctact cgggacccct taccgaaaca tcgccgcatt ctgcagagga gtcgagtgta
     2641 tgtaaacttc tgacccactg ggaatgtgat gaaagaaata aaagctgaaa tgaatcattc
     2701 tctctactat tattctgata tttcacattc ttaaaataaa gtggtgatcc taactgacct
     2761 aagacaggga atttttacta ggattaaatg tcaggaattg tgaaaaagtg agtttaaatg
     2821 tatttggcta aggtgtatgt aaacttccga cttcaactgt atagggatct gcttcctcgc
     2881 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2941 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3001 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3061 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3121 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3181 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3241 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3301 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3361 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3421 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3481 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3541 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3601 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3661 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3721 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3781 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3841 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3901 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3961 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4021 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4081 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4141 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4201 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4261 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4321 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4381 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4441 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4501 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4561 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4621 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4681 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4741 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgatg
     4801 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     4861 ttaagatccc tata
//
