LOCUS       sequence-372481-        8388 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 8388)
  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 8388)
  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-NTERM-GFP
FEATURES             Location/Qualifiers
     source          1..8388
                     /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             3251..3292
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(3536..3538,3539..4249)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    4331..4448
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4527..5115
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4580..4600)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4998..5009)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5187..5420
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5498..5619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5535..5554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5589..5608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5659..5794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5814..5833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5815..5833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5843..5860)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5843..5859)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5852..5874)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6001..6456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6088..6107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6298..6319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6482..6586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6587..6655,6656..7447)
                     /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(6805..6824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7618..8206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8107..8126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8360..8377
                     /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 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgttcaatgt
     3301 cttaatggtt cataagcgaa gccatactgg tgaacgccca ttccagtgta atcagtgtgg
     3361 ggcatctttt actcagaaag gtaacctcct ccgccacatt aaactgcaca caggggaaaa
     3421 accttttaag tgtcacctct gcaactatgc atgccaaaga agagatgcgc tcacgcgtgc
     3481 tgaagctgct gcaaaggaag ctgcagctaa ggaggctgca gctaaggctg gatccgtcag
     3541 taaaggggaa gaattgttta cgggtgttgt tccgatactt gttgaattgg atggggatgt
     3601 gaatggtcat aaattttccg ttagtggtga aggggaaggt gacgctactt atggtaaatt
     3661 gacattgaaa tttatttgta ctacaggaaa acttccagtc ccatggccga ctttggtaac
     3721 gactcttaca tatggagtcc aatgttttag taggtatcca gatcacatga aacaacatga
     3781 tttctttaaa tcagcaatgc ctgagggata tgttcaagaa cgaacaatat tctttaaaga
     3841 tgatgggaac tataagacaa gggcagaagt caaatttgaa ggtgatacac tcgttaatcg
     3901 gattgaactg aaagggatag attttaagga agatgggaat attcttgggc acaaacttga
     3961 atataattat aattcccata atgtatacat aatggctgac aagcagaaga atggaattaa
     4021 agtcaatttt aaaataaggc ataatataga agatgggagt gtccaattgg ctgatcatta
     4081 ccaacaaaat acacctattg gggatggtcc tgtcctcttg cctgataatc attatctttc
     4141 tacacaatct gctctctcaa aggaccctaa tgaaaagcgc gaccatatgg tgttgttgga
     4201 atttgtcact gctgcaggca ttaccctggg aatggatgaa ctctataaat gagaattctg
     4261 acggagtaca agtaaactag taagcttggc gtaactagat cttgagacaa atggcagtat
     4321 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     4381 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     4441 aaaattttcg ggtttattac agggacagca gagatccact ttgggctcga gcttggttca
     4501 ttctcaagcc tacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     4561 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4621 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4681 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4741 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4801 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     4861 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     4921 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     4981 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5041 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5101 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     5161 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5221 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5281 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5341 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5401 agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt
     5461 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     5521 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5581 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     5641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5761 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     5821 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5881 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5941 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     6001 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6061 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     6481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     8281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     8341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
