LOCUS       sequence-372473-        8284 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 8284)
  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 8284)
  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-NTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8284
                     /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             join(3436..3438,3439..4149)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    4227..4344
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4423..5011
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4476..4496)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4894..4905)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5083..5316
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5394..5515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5431..5450)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5485..5504
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5555..5690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5710..5729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5711..5729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5739..5756)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5739..5755)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5748..5770)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5897..6352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5984..6003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6194..6215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6378..6482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6483..6551,6552..7343)
                     /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(6701..6720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7514..8102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8003..8022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8256..8273
                     /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 cgttcaatgt cttaatggtt cataagcgaa gccatactgg tgaacgccca
     3241 ttccagtgta atcagtgtgg ggcatctttt actcagaaag gtaacctcct ccgccacatt
     3301 aaactgcaca caggggaaaa accttttaag tgtcacctct gcaactatgc atgccaaaga
     3361 agagatgcgc tcacgcgtgc tgaagctgct gcaaaggaag ctgcagctaa ggaggctgca
     3421 gctaaggctg gatccgtcag taaaggggaa gaattgttta cgggtgttgt tccgatactt
     3481 gttgaattgg atggggatgt gaatggtcat aaattttccg ttagtggtga aggggaaggt
     3541 gacgctactt atggtaaatt gacattgaaa tttatttgta ctacaggaaa acttccagtc
     3601 ccatggccga ctttggtaac gactcttaca tatggagtcc aatgttttag taggtatcca
     3661 gatcacatga aacaacatga tttctttaaa tcagcaatgc ctgagggata tgttcaagaa
     3721 cgaacaatat tctttaaaga tgatgggaac tataagacaa gggcagaagt caaatttgaa
     3781 ggtgatacac tcgttaatcg gattgaactg aaagggatag attttaagga agatgggaat
     3841 attcttgggc acaaacttga atataattat aattcccata atgtatacat aatggctgac
     3901 aagcagaaga atggaattaa agtcaatttt aaaataaggc ataatataga agatgggagt
     3961 gtccaattgg ctgatcatta ccaacaaaat acacctattg gggatggtcc tgtcctcttg
     4021 cctgataatc attatctttc tacacaatct gctctctcaa aggaccctaa tgaaaagcgc
     4081 gaccatatgg tgttgttgga atttgtcact gctgcaggca ttaccctggg aatggatgaa
     4141 ctctataaat gagaattctg agtacaagta aactagtaag cttggcgtaa ctagatcttg
     4201 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     4261 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     4321 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     4381 gctcgagctt ggttcattct caagcctacg cgttaagtcg acaatcaacc tctggattac
     4441 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4501 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4561 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4621 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4681 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4741 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4801 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     4861 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4921 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4981 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     5041 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5101 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5161 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5221 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5281 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     5341 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5521 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     5701 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     5761 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5821 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5881 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6301 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     6361 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6421 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6481 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6541 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6601 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6661 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6721 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6781 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6841 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6901 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6961 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7021 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7081 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7141 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7201 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7261 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7321 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7381 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7441 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7501 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7621 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7681 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7741 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7801 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7861 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7921 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7981 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8041 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8101 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8161 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8221 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8281 aaga
//
