LOCUS       sequence-372500-        8299 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 8299)
  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 8299)
  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-CTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8299
                     /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             join(3157..3159,3160..3870)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             3928..4131
                     /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             4132..4173
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4242..4359
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4438..5026
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4491..4511)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4909..4920)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5098..5331
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5409..5530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5500..5519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5570..5705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5725..5744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5726..5744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5754..5771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5754..5770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5763..5785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5912..6367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5999..6018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6209..6230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6393..6497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6498..6566,6567..7358)
                     /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(6716..6735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7529..8117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8018..8037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8271..8288
                     /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 ggatccgtca gtaaagggga agaattgttt
     3181 acgggtgttg ttccgatact tgttgaattg gatggggatg tgaatggtca taaattttcc
     3241 gttagtggtg aaggggaagg tgacgctact tatggtaaat tgacattgaa atttatttgt
     3301 actacaggaa aacttccagt cccatggccg actttggtaa cgactcttac atatggagtc
     3361 caatgtttta gtaggtatcc agatcacatg aaacaacatg atttctttaa atcagcaatg
     3421 cctgagggat atgttcaaga acgaacaata ttctttaaag atgatgggaa ctataagaca
     3481 agggcagaag tcaaatttga aggtgataca ctcgttaatc ggattgaact gaaagggata
     3541 gattttaagg aagatgggaa tattcttggg cacaaacttg aatataatta taattcccat
     3601 aatgtataca taatggctga caagcagaag aatggaatta aagtcaattt taaaataagg
     3661 cataatatag aagatgggag tgtccaattg gctgatcatt accaacaaaa tacacctatt
     3721 ggggatggtc ctgtcctctt gcctgataat cattatcttt ctacacaatc tgctctctca
     3781 aaggacccta atgaaaagcg cgaccatatg gtgttgttgg aatttgtcac tgctgcaggc
     3841 attaccctgg gaatggatga actctataaa gaattcgctg aagctgctgc aaaggaagct
     3901 gcagctaagg aggctgcagc taaggctaag gagaagagtg cttgtcctaa agatccagcc
     3961 aaacctccgg ccaaggcaca agttgtggga tggccaccgg tgagatcata ccggaagaac
     4021 gtgatggttt cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt cgtgaaggta
     4081 tcaatggacg gagcaccgta cttgaggaaa atcgatttga ggatgtataa aggtaagcct
     4141 atccctaacc ctctcctcgg tctcgattct acgtgagtac aagtaaacta gtaagcttgg
     4201 cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     4261 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     4321 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     4381 agagatccac tttgggctcg agcttggttc attctcaagc ctacgcgtta agtcgacaat
     4441 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4501 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4561 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4621 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4681 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4741 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4801 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4861 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4921 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4981 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     5041 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5101 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     5161 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5221 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5281 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     5341 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     5401 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5521 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     5581 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5641 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5701 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     5761 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     5821 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5881 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5941 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6001 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6061 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6121 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6241 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     6361 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6421 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6481 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6541 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6601 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6661 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6721 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6781 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6841 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6901 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6961 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7021 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7081 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7141 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7201 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7261 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7321 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7381 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7441 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7501 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7561 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7621 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7681 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7741 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7801 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7861 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7921 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7981 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8041 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8101 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8161 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     8221 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     8281 tgagcgagga agcggaaga
//
