LOCUS       sequence-372501-        8302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Degron-GFP fusion plasmid with restriction sites BamH1 and EcoR1 
            around GFP for subcloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8302)
  AUTHORS   Bondeson DP, Mullin-Bernstein Z, Oliver S, Skipper TA, Atack TC, 
            Bick N, Ching M, Guirguis AA, Kwon J, Langan C, Millson D, Paolella 
            BR, Tran K, Wie SJ, Vazquez F, Tothova Z, Golub TR, Sellers WR, 
            Ianari A
  TITLE     Systematic profiling of conditional degron tag technologies for 
            target validation studies.
  JOURNAL   Nat Commun. 2022 Sep 20;13(1):5495. doi: 10.1038/s41467-022-33246-4.
  PUBMED    36127368
REFERENCE   2  (bases 1 to 8302)
  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: "Nat 
            Commun."; date: "2022-09-20"; volume: "13(1):5495. doi"; pages: " 
            10.1038/s41467-022-33246-4"
COMMENT     Sequence Label: SFFV-AID-NTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     239..259
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        241..259
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        287..513
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             514..694
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    741..866
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1359..1592
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1777..1821
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2023..2429
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3094..3150
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3151..3192
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3193..3396
                     /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"
     CDS             join(3454..3456,3457..4167)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    4245..4362
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4441..5029
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4494..4514)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4912..4923)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5101..5334
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5412..5533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5449..5468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5503..5522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5573..5708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5728..5747)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5729..5747)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5757..5774)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5757..5773)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5766..5788)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5915..6370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6002..6021)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6212..6233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6396..6500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6501..6569,6570..7361)
                     /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     complement(6719..6738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7532..8120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8021..8040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8274..8291
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
      241 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca
      301 atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag
      361 aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga
      421 aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat
      481 attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga
      541 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      601 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      661 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      721 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      781 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      841 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      901 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      961 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1021 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1081 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1141 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1201 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1261 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1321 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1381 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1441 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1501 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1561 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1621 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1681 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1741 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1801 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1861 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1921 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1981 accccgaggg gaccctattc cagcacatat gagctagctg cagtaacgcc attttgcaag
     2041 gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa
     2101 tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc
     2161 caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc
     2221 agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca
     2281 aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct
     2341 gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg
     2401 ccagtcctcc gacagactga gtcggccggt cgaatcaagc ttatcgatac cgtcgactcc
     2461 ggaatagcca ccatagccac cgccaccatg accgagtaca agccaactgt ccgcttggca
     2521 acccgcgacg atgtgcccag ggccgtccgc acgctggctg ctgccttcgc cgactaccca
     2581 gccacccgcc acaccgtgga cccggaccgc cacatcgagc gcgtcaccga gctgcaggag
     2641 ctcttcctca cccgagttgg ccttgacatt ggcaaggtct gggtggcgga cgacggtgct
     2701 gccgtggccg tctggaccac ccccgagtcg gtcgaggctg gcgccgtgtt cgccgagatc
     2761 gggccgcgca tggccgagct ttcgggctcc cggttggctg cccaacagca gatggagggg
     2821 ctgttggcac cccaccgccc caaggagccg gcctggttcc tggccacagt tggtgtctct
     2881 cccgaccacc agggcaaggg cctgggcagc gccgtggtgc tgcctggggt cgaggctgcc
     2941 gagcgcgctg gcgtccctgc cttcctggag acctctgccc ctcgcaactt gcccttctac
     3001 gagcgcctgg gcttcaccgt caccgcggac gtcgaggtgc ccgaggggcc gcgcacctgg
     3061 tgcatgaccc gcaagcctgg ggcaggctcc ggagccacca acttttccct gctgaagcag
     3121 gctggcgacg tcgaggagaa ccctggccct ggtaagccta tccctaaccc tctcctcggt
     3181 ctcgattcta cgaaggagaa gagtgcttgt cctaaagatc cagccaaacc tccggccaag
     3241 gcacaagttg tgggatggcc accggtgaga tcataccgga agaacgtgat ggtttcctgc
     3301 caaaaatcaa gcggtggccc ggaggcggcg gcgttcgtga aggtatcaat ggacggagca
     3361 ccgtacttga ggaaaatcga tttgaggatg tataaagctg aagctgctgc aaaggaagct
     3421 gcagctaagg aggctgcagc taaggctgga tccgtcagta aaggggaaga attgtttacg
     3481 ggtgttgttc cgatacttgt tgaattggat ggggatgtga atggtcataa attttccgtt
     3541 agtggtgaag gggaaggtga cgctacttat ggtaaattga cattgaaatt tatttgtact
     3601 acaggaaaac ttccagtccc atggccgact ttggtaacga ctcttacata tggagtccaa
     3661 tgttttagta ggtatccaga tcacatgaaa caacatgatt tctttaaatc agcaatgcct
     3721 gagggatatg ttcaagaacg aacaatattc tttaaagatg atgggaacta taagacaagg
     3781 gcagaagtca aatttgaagg tgatacactc gttaatcgga ttgaactgaa agggatagat
     3841 tttaaggaag atgggaatat tcttgggcac aaacttgaat ataattataa ttcccataat
     3901 gtatacataa tggctgacaa gcagaagaat ggaattaaag tcaattttaa aataaggcat
     3961 aatatagaag atgggagtgt ccaattggct gatcattacc aacaaaatac acctattggg
     4021 gatggtcctg tcctcttgcc tgataatcat tatctttcta cacaatctgc tctctcaaag
     4081 gaccctaatg aaaagcgcga ccatatggtg ttgttggaat ttgtcactgc tgcaggcatt
     4141 accctgggaa tggatgaact ctataaatga gaattctagg tacaagtaaa ctagtaagct
     4201 tggcgtaact agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     4261 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     4321 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     4381 agcagagatc cactttgggc tcgagcttgg ttcattctca agcctacgcg ttaagtcgac
     4441 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4501 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4561 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4621 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4681 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4741 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4801 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4861 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4921 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4981 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     5041 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5101 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     5161 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5221 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5281 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
     5341 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     5401 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5521 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     5581 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5641 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5701 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     5761 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5821 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5881 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     5941 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6001 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6061 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6121 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6241 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     6301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6361 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6421 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6481 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6541 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6601 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6661 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6721 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6781 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6841 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6901 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6961 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7021 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7081 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7141 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7201 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7261 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7321 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8281 cagtgagcga ggaagcggaa ga
//
