LOCUS       sequence-372468-        8378 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 8378)
  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 8378)
  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: PGK-IKZF3-CTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8378
                     /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        2012..2512
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2431..2449
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3194..3250
                     /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(3257..3259,3260..3970)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             4208..4249
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4321..4438
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4517..5105
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4570..4590)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4988..4999)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5177..5410
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5488..5609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5525..5544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5579..5598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5649..5784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5804..5823)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5805..5823)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5833..5850)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5833..5849)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5842..5864)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5991..6446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6078..6097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6288..6309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6472..6576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6577..6645,6646..7437)
                     /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(6795..6814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7608..8196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8097..8116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8350..8367
                     /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 gttgcgcctt ttccaaggca gccctgggtt
     2041 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
     2101 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
     2161 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
     2221 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
     2281 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
     2341 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     2401 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
     2461 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agcagactga
     2521 gtcggccggt cgaatcaagc ttatcgatac cgtcgactcc ggaatagcca ccatagccac
     2581 cgccaccatg accgagtaca agccaactgt ccgcttggca acccgcgacg atgtgcccag
     2641 ggccgtccgc acgctggctg ctgccttcgc cgactaccca gccacccgcc acaccgtgga
     2701 cccggaccgc cacatcgagc gcgtcaccga gctgcaggag ctcttcctca cccgagttgg
     2761 ccttgacatt ggcaaggtct gggtggcgga cgacggtgct gccgtggccg tctggaccac
     2821 ccccgagtcg gtcgaggctg gcgccgtgtt cgccgagatc gggccgcgca tggccgagct
     2881 ttcgggctcc cggttggctg cccaacagca gatggagggg ctgttggcac cccaccgccc
     2941 caaggagccg gcctggttcc tggccacagt tggtgtctct cccgaccacc agggcaaggg
     3001 cctgggcagc gccgtggtgc tgcctggggt cgaggctgcc gagcgcgctg gcgtccctgc
     3061 cttcctggag acctctgccc ctcgcaactt gcccttctac gagcgcctgg gcttcaccgt
     3121 caccgcggac gtcgaggtgc ccgaggggcc gcgcacctgg tgcatgaccc gcaagcctgg
     3181 ggcaggctcc ggagccacca acttttccct gctgaagcag gctggcgacg tcgaggagaa
     3241 ccctggccct ggatccgtca gtaaagggga agaattgttt acgggtgttg ttccgatact
     3301 tgttgaattg gatggggatg tgaatggtca taaattttcc gttagtggtg aaggggaagg
     3361 tgacgctact tatggtaaat tgacattgaa atttatttgt actacaggaa aacttccagt
     3421 cccatggccg actttggtaa cgactcttac atatggagtc caatgtttta gtaggtatcc
     3481 agatcacatg aaacaacatg atttctttaa atcagcaatg cctgagggat atgttcaaga
     3541 acgaacaata ttctttaaag atgatgggaa ctataagaca agggcagaag tcaaatttga
     3601 aggtgataca ctcgttaatc ggattgaact gaaagggata gattttaagg aagatgggaa
     3661 tattcttggg cacaaacttg aatataatta taattcccat aatgtataca taatggctga
     3721 caagcagaag aatggaatta aagtcaattt taaaataagg cataatatag aagatgggag
     3781 tgtccaattg gctgatcatt accaacaaaa tacacctatt ggggatggtc ctgtcctctt
     3841 gcctgataat cattatcttt ctacacaatc tgctctctca aaggacccta atgaaaagcg
     3901 cgaccatatg gtgttgttgg aatttgtcac tgctgcaggc attaccctgg gaatggatga
     3961 actctataaa gaattcgctg aagctgctgc aaaggaagct gcagctaagg aggctgcagc
     4021 taaggctttc aatgtcttaa tggttcataa gcgaagccat actggtgaac gcccattcca
     4081 gtgtaatcag tgtggggcat cttttactca gaaaggtaac ctcctccgcc acattaaact
     4141 gcacacaggg gaaaaacctt ttaagtgtca cctctgcaac tatgcatgcc aaagaagaga
     4201 tgcgctcggt aagcctatcc ctaaccctct cctcggtctc gattctacgt agacgcgtgt
     4261 acaagtaaac taagcttggc gtaactagat cttgagacaa atggcagtat tcatccacaa
     4321 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     4381 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     4441 ggtttattac agggacagca gagatccact ttgggctcga gcttggttca ttctcaagcc
     4501 tacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4561 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4621 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4681 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4741 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4801 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4861 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4921 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4981 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5041 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5101 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5161 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     5221 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5281 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5341 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5401 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     5461 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     5521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5581 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     5641 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5701 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5761 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     5821 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5881 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5941 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     6001 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6061 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6121 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6181 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6241 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6301 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6361 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6421 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     6481 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6541 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6601 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6661 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6721 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6781 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6841 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6901 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6961 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7021 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7081 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7141 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7201 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7261 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7321 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7381 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7441 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7501 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7621 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7681 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7741 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7801 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7861 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7921 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7981 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8041 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8101 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8161 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8221 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8281 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8341 accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
