LOCUS       sequence-234361-        7256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retrovirus vector expresses AID-fusion protein under TRE promoter 
            and with hygromycin resistance selectio.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  AUTHORS   Ng LY, Ma HT, Liu JCY, Huang X, Lee N, Poon RYC
  TITLE     Conditional gene inactivation by combining tetracycline-mediated 
            transcriptional repression and auxin-inducible degron-mediated 
            degradation.
  JOURNAL   Cell Cycle. 2018 Dec 23. doi: 10.1080/15384101.2018.1563395.
  PUBMED    30582405
REFERENCE   2  (bases 1 to 7256)
  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: "Cell Cycle.
            2018 Dec 23. doi: 10.1080/15384101.2018.1563395."
COMMENT     Sequence Label: pREVTRE-AID
FEATURES             Location/Qualifiers
     source          1..7256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             440..615
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    678..877
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1078..1494
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1424..1446
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1462..1478
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1551..2570
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     protein_bind    join(2895..2913,2914..2936,2937..2955,2956..2978,2979..2997,
                     2998..3020,3021..3039,3040..3062,3063..3081,3082..3104,
                     3105..3123,3124..3146,3147..3165)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3198..3236
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3233..3257
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3332..4015
                     /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"
     LTR             4184..4777
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4880..4902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4988..5128
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5043..5062
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5143..5160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5314..5902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5394..5413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6073..6864,6865..6933))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6696..6715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6934..7038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7106..7124
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agccgcggcc cttccgtttc tttgcttttg aaagacccca cccgtaggtg gcaagctagc
       61 ttaagtaacg ccactttgca aggcatggaa aaatacataa ctgagaatag aaaagttcag
      121 atcaaggtca ggaacaaaga aacagctgaa taccaaacag gatatctgtg gtaagcggtt
      181 cctgccccgg ctcagggcca agaacagatg agacagctga gtgatgggcc aaacaggata
      241 tctgtggtaa gcagttcctg ccccggctcg gggccaagaa cagatggtcc ccagatgcgg
      301 tccagccctc agcagtttct agtgaatcat cagatgtttc cagggtgccc caaggacctg
      361 aaaatgaccc tgtaccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttccgct ctccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtct
      481 tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat
      541 ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccacga
      601 cgggggtctt tcatttgggg gctcgtccgg gatttggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgtttg atgttatgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc
      841 agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg
      901 accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
      961 tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc
     1021 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc
     1081 cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc
     1141 gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa
     1201 tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc
     1261 caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac
     1321 atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac
     1381 cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt
     1441 tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt
     1501 gcgatctgat cagcttgcca caacccgtac caaagatgga tagatccgga aagcctgaac
     1561 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     1621 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     1681 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     1741 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga
     1801 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     1861 ccgaactgcc cgctgttctg cagccggtcg cggaggctat ggatgcgatc gctgcggccg
     1921 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     1981 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     2041 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     2101 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     2161 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     2221 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     2281 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     2341 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     2401 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     2461 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     2521 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag agtagatgcc
     2581 gaccgaacaa gagctgattt cgagaacgcc tcagccagca actcgcgcga gcctagcaag
     2641 gcaaatgcga gagaacggcc ttacgcttgg tggcacagtt ctcgtccaca gttcgctaag
     2701 ctcgctcggc tgggtcgcgg gagggccggt cgcagtgatt caggcccttc tggattgtgt
     2761 tggtccccag ggcacgattg tcatgcccac gcactcgggt gatctgactg atcccgcaga
     2821 ttggagatcg ccgcccgtgc ctgccgattg ggtgcagatc tcgacgccct ttcgtcttta
     2881 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2941 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     3001 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     3061 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     3121 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     3181 ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
     3241 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagag
     3301 accgggaccg atccagcctc cgcgggctag catgggcagt gtcgagctga atctgaggga
     3361 gactgagctg tgtcttggtc ttcccggtgg agatacagtg gctccggtaa ccggaaacaa
     3421 gagagggttc tcagagacgg ttgatctgaa gctaaatctg aataatgagc ctgcaaacaa
     3481 ggaaggatct acgactcatg acgtcgtgac ttttgattcc aaggagaaga gtgcttgtcc
     3541 taaagatcca gccaaacctc cggccaaggc acaagttgtg ggatggccac cggtgagatc
     3601 ataccggaag aacgtgatgg tttcctgcca aaaatcaagc ggtggcccgg aggcggcggc
     3661 gttcgtgaag gtatcaatgg acggagcacc gtacttgagg aaaatcgatt tgaggatgta
     3721 taaaagctac gatgagcttt ctaatgcttt gtccaacatg ttcagctctt ttacgatggg
     3781 caaacatgga ggagaagaag gaatgataga cttcatgaat gagaggaaat tgatggattt
     3841 ggtgaatagc tgggactatg ttccctctta tgaagacaaa gacggtgatt ggatgctcgt
     3901 cggcgacgtt ccttggccaa tgttcgtcga tacatgcaag cgtttacgtc tcatgaaagg
     3961 atcggatgcc attggtctcg ctccgagggc gatggagaag tgcaagagca gagctgcggc
     4021 cggagctggt gcaggcgctg gagcgggtgc cgatatccta ttcaccatgg ctcgagaatt
     4081 cgagctcggt acccggggat ccgtcgaggc cgcctcggcc aagcttgcgg ccgcaaacat
     4141 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4201 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     4261 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     4321 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     4381 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4441 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     4501 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4561 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4621 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4681 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4741 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     4801 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     4861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4981 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     5041 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5101 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     5161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6361 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     6421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6661 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     6721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7081 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     7141 attgatacca gatcaccgaa aactgtcctc caaatgtgtc cccctcacac tcccaaattc
     7201 gcgggcttct gcctcttaga ccactctacc ctattcccca cactcaccgg agccaa
//
