LOCUS       sequence-338411-        4835 bp    DNA     circular SYN 14-APR-2025
DEFINITION  TRE-controlled mAID-fusion sleeping beauty transposon vector with 
            selection marker (zeocin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  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 4835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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/Zeo
FEATURES             Location/Qualifiers
     source          1..4835
                     /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..2244
                     /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
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(2281..2298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2287..2511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2837..2854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3008..3596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3088..3107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3767..4558,4559..4627))
                     /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     4390..4409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4628..4732)
                     /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 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     1921 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     1981 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     2041 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     2101 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     2161 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     2221 ttcgtggccg aggagcagga ctgaagcttg gggatcaatt ctctagagct cgctgatcag
     2281 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2341 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2401 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2461 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2521 cggaaagaac cagctgggcc tactagctac tcgggacccc ttaccgaaac atcgccgcat
     2581 tctgcagagg agtcgagtgt atgtaaactt ctgacccact gggaatgtga tgaaagaaat
     2641 aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt cttaaaataa
     2701 agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat gtcaggaatt
     2761 gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg acttcaactg
     2821 tatagggatc tgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2881 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2941 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3421 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3541 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3661 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3721 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4741 cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4801 cgcatcagga aattgtaagc gttaagatcc ctata
//
