LOCUS       sequence-231816-        6767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminus of plum pox virus protease (PPVp) fused to the 
            ABI-abscisic acid responsive protein domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6767)
  AUTHORS   Fink T, Lonzaric J, Praznik A, Plaper T, Merljak E, Leben K, Jerala 
            N, Lebar T, Strmsek Z, Lapenta F, Bencina M, Jerala R
  TITLE     Design of fast proteolysis-based signaling and logic circuits in 
            mammalian cells.
  JOURNAL   Nat Chem Biol. 2019 Feb;15(2):115-122. doi: 
            10.1038/s41589-018-0181-6. Epub 2018 Dec 10.
  PUBMED    30531965
REFERENCE   2  (bases 1 to 6767)
  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"; doi: 
            "10.1038/s41589-018-0181-6"; journalName: "Nat Chem Biol"; date: 
            "2019-02"; volume: "15"; issue: "2"; pages: "115-122"
COMMENT     Sequence Label: ABI_cPPVp
FEATURES             Location/Qualifiers
     source          1..6767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             953..982
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2320..2338)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2321..2338)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2358..2375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2364..2588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2634..3062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2721..2740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2931..2952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3071..3091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3076..3405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3256..3391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3318..3337
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3472..4266
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3526..3545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4136..4155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4440..4561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4477..4496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4531..4550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4610..4626)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4610..4626)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4623..4645)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4634..4650
                     /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)."
     promoter        complement(join(4658..4664,4665..4682,4683..4688))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4703..4724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4841..4858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5012..5600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5092..5111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5771..6562,6563..6631))
                     /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     6394..6413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6632..6736)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tggagcagaa
      961 gctgatcagc gaggaggatc tgacgcgtgt gcctttgtat ggttttactt cgatttgtgg
     1021 aagaagacct gagatggaag ctgctgtttc gactatacca agattccttc aatcttcctc
     1081 tggttcgatg ttagatggtc ggtttgatcc tcaatccgcc gctcatttct tcggtgttta
     1141 cgacggccat ggcggttctc aggtagcgaa ctattgtaga gagaggatgc atttggcttt
     1201 ggcggaggag atagctaagg agaaaccgat gctctgcgat ggtgatacgt ggctggagaa
     1261 gtggaagaaa gctcttttca actcgttcct gagagttgac tcggagattg agtcagttgc
     1321 gccggagacg gttgggtcaa cgtcggtggt tgccgttgtt ttcccgtctc acatcttcgt
     1381 cgctaactgc ggtgactcta gagccgttct ttgccgcggc aaaactgcac ttccattatc
     1441 cgttgaccat aaaccggata gagaagatga agctgcgagg attgaagccg caggagggaa
     1501 agtgattcag tggaatggag ctcgtgtttt cggtgttctc gccatgtcga gatccattgg
     1561 cgatagatac ttgaaaccat ccatcattcc tgatccggaa gtgacggctg tgaagagagt
     1621 aaaagaagat gattgtctga ttttggcgag tgacggggtt tgggatgtaa tgacggatga
     1681 agaagcgtgt gagatggcaa ggaagcggat tctcttgtgg cacaagaaaa acgcggtggc
     1741 tggggatgca tcgttgctcg cggatgagcg gagaaaggaa gggaaagatc ctgcggcgat
     1801 gtccgcggct gagtatttgt caaagctggc gatacagaga ggaagcaaag acaacataag
     1861 tgtggtggtg gttgatttga agggatccgg aagtaagagc atcagcagca ccatgagcga
     1921 gaccagcgcc acctaccccg tggacaacag ccacttctgg aagcactgga tcagcaccaa
     1981 ggacggccac tgcggcctgc ccatcgtgag cacccgcgac ggcagcatcc tgggcctgca
     2041 cagcctggcc aacagcacca acacccagaa cttctacgcc gccttccccg acaacttcga
     2101 gaccacctac ctgagcaacc aggacaacga caactggatc aagcagtggc gctacaaccc
     2161 cgacgaggtg tgctggggca gcctgcaact gaagcgcgac atcccccaga gccccttcac
     2221 catctgcaag ctgctgaccg acctggacgg cgagttcgtg tacacccagt aaactagtgc
     2281 ggccgcctgc agctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     2341 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2401 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2461 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2521 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2581 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3061 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3421 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3781 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4261 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
