LOCUS       sequence-338221-        6026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-controlled AID-fusion sleeping beauty transposon vector with 
            selection marker (hygromycin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  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 6026)
  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/Hyg
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..770
                     /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    884..1005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(895..914)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     949..968
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1737..2756
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     primer_bind     complement(2811..2828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2817..3041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3367..3384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3538..4126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3618..3637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4297..5088,5089..5157))
                     /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     4920..4939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5158..5262)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    join(5676..5694,5695..5717,5718..5736,5737..5759,5760..5778,
                     5779..5801,5802..5820,5821..5843,5844..5862,5863..5885,
                     5886..5904,5905..5927,5928..5946)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        5979..6017
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6014..6026,1..12)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagagaccgg
       61 gaccgatcca gcctccgcgg gctagcatgg gcagtgtcga gctgaatctg agggagactg
      121 agctgtgtct tggtcttccc ggtggagata cagtggctcc ggtaaccgga aacaagagag
      181 ggttctcaga gacggttgat ctgaagctaa atctgaataa tgagcctgca aacaaggaag
      241 gatctacgac tcatgacgtc gtgacttttg attccaagga gaagagtgct tgtcctaaag
      301 atccagccaa acctccggcc aaggcacaag ttgtgggatg gccaccggtg agatcatacc
      361 ggaagaacgt gatggtttcc tgccaaaaat caagcggtgg cccggaggcg gcggcgttcg
      421 tgaaggtatc aatggacgga gcaccgtact tgaggaaaat cgatttgagg atgtataaaa
      481 gctacgatga gctttctaat gctttgtcca acatgttcag ctcttttacg atgggcaaac
      541 atggaggaga agaaggaatg atagacttca tgaatgagag gaaattgatg gatttggtga
      601 atagctggga ctatgttccc tcttatgaag acaaagacgg tgattggatg ctcgtcggcg
      661 acgttccttg gccaatgttc gtcgatacat gcaagcgttt acgtctcatg aaaggatcgg
      721 atgccattgg tctcgctccg agggcgatgg agaagtgcaa gagcagagct gcggccggag
      781 ctggtgcagg cgctggagcg ggtgccgata tcctattcac catggctcga gaattcgagc
      841 tcggtacccg gggatccaga catgataaga tcgatagaca tgataagata cattgatgag
      901 tttggacaaa ccacaacaag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
      961 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1021 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     1081 ctctacaaat gtggtaccta gacgagaccc tgtctcacaa aataaagtaa gcccggactg
     1141 agtgcggaaa ggcgggcctg gcgggtctgg tctccccatg cgggccacca gaggccctgc
     1201 agccttcagt cgcttgaagg ggtaatggcg cttccactca caaacatggc ggacagagcg
     1261 tgtgaacgag atgaacagcc cctcaaaaat atggccgccg aggctggacg gccgtgcccc
     1321 agcagcaccg cctccgcgcc ccacgtgatc tctcgccggg cacagcgctg accgcggagg
     1381 tccaaccgga agaatgtccg gattggacat tcggaagagg gcccgccttc cctggggaat
     1441 ctctgcgcac gcgcagaacg cttcgaccaa tgaaaacaca ggaagccgtc cgcgcaaccg
     1501 cgttgcgtca cttctgccgc ccctgtttca aggtatatag ccgtagacgg aacttcgcct
     1561 ttctctcggc cttagcgcca tttttttggg tgagtgtttt ttggttcctg cgttgggatt
     1621 ccgtgtacaa tccatagaca tctgacctcg gcacttagca tcatcacagc aaactaactg
     1681 tagcctttct ctctttccct gtagaaacct ctgcactcga catggataga tccggaaagc
     1741 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     1801 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     1861 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     1921 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttca
     1981 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     2041 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggctatggat gcgatcgctg
     2101 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     2161 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     2221 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     2281 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     2341 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     2401 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     2461 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     2521 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     2581 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     2641 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     2701 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagttcg
     2761 aaggcctgtc gtgaagcttg gggatcaatt ctctagagct cgctgatcag cctcgactgt
     2821 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2881 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2941 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3001 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3061 cagctgggcc tactagctac tcgggacccc ttaccgaaac atcgccgcat tctgcagagg
     3121 agtcgagtgt atgtaaactt ctgacccact gggaatgtga tgaaagaaat aaaagctgaa
     3181 atgaatcatt ctctctacta ttattctgat atttcacatt cttaaaataa agtggtgatc
     3241 ctaactgacc taagacaggg aatttttact aggattaaat gtcaggaatt gtgaaaaagt
     3301 gagtttaaat gtatttggct aaggtgtatg taaacttccg acttcaactg tatagggatc
     3361 tgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3421 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3481 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3541 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3601 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3661 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3721 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3781 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3841 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3901 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3961 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4021 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4081 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4141 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4201 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4261 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4321 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4381 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4441 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4501 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4561 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4621 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4681 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4741 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4801 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4861 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4921 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4981 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5041 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5101 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5161 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5221 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5281 gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     5341 aattgtaagc gttaagatcc ctatacagtt gaagtcggaa gtttacatac acttaagttg
     5401 gagtcattaa aactcgtttt tcaactactc cacaaatttc ttgttaacaa acaatagttt
     5461 tggcaagtca gttaggacat ctactttgtg catgacacaa gtcatttttc caacaattgt
     5521 ttacagacag attatttcac ttataattca ctgtatcaca attccagtgg gtcagaagtt
     5581 tacatacact aagttcgact cctctgcaga atgcggcgat gtttcggtaa ggggtccgct
     5641 atctaggccc tttcgtcttt agtcgagttt accactccct atcagtgata gagaaaagtg
     5701 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     5761 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     5821 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     5881 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     5941 tagagaaaag tgaaagtcga gctcggtacc cgggtcgagg taggcgtgta cggtgggagg
     6001 cctatataag cagagctcgt ttagtg
//
