LOCUS       sequence-372486-        8275 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 8275)
  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 8275)
  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-IKZF3-CTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8275
                     /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             4108..4149
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4218..4335
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4414..5002
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4467..4487)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4885..4896)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5074..5307
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5385..5506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5422..5441)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5476..5495
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5546..5681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5701..5720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5702..5720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5730..5747)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5730..5746)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5739..5761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5888..6343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5975..5994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6185..6206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6369..6473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6474..6542,6543..7334)
                     /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(6692..6711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7505..8093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7994..8013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8247..8264
                     /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 taaggctttc aatgtcttaa tggttcataa gcgaagccat
     3961 actggtgaac gcccattcca gtgtaatcag tgtggggcat cttttactca gaaaggtaac
     4021 ctcctccgcc acattaaact gcacacaggg gaaaaacctt ttaagtgtca cctctgcaac
     4081 tatgcatgcc aaagaagaga tgcgctcggt aagcctatcc ctaaccctct cctcggtctc
     4141 gattctacgt aggtacaagt aaactagtaa gcttggcgta actagatctt gagacaaatg
     4201 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     4261 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     4321 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg ggctcgagct
     4381 tggttcattc tcaagcctac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     4441 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4501 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4561 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4621 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4681 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4741 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4801 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4861 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4921 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4981 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     5041 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5101 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5161 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5221 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5281 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     5341 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     5401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     5521 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5581 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5641 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     5701 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5761 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5821 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5881 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5941 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6001 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6061 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6121 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6181 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6241 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6301 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     6361 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6421 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6481 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6541 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6601 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6661 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6721 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6781 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6841 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6901 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6961 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7021 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7081 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7141 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7201 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7261 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7321 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7381 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7441 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7501 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7561 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7621 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7681 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7741 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7801 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7861 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7921 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7981 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8041 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8101 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8161 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8221 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaga
//
