LOCUS       sequence-369576-        7314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin regulated PTPPest with mVenus tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  AUTHORS   Fauser J, Huyot V, Matsche J, Szynal BN, Alexeev Y, Kota P, Karginov
            AV
  TITLE     Dissecting protein tyrosine phosphatase signaling by engineered 
            chemogenetic control of its activity.
  JOURNAL   J Cell Biol. 2022 Aug 1;221(8). pii: 213352. doi: 
            10.1083/jcb.202111066. Epub 2022 Jul 13.
  PUBMED    35829702
REFERENCE   2  (bases 1 to 7314)
  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: "J Cell 
            Biol."; date: "2022-08-1"; volume: "221(8). pii: 213352. doi"; 
            pages: " 10.1083/jcb.202111066. Epub 2022 Jul 13"
COMMENT     Sequence Label: mVenus-RapR-PTPPest
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4102..4223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4230..4685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4818..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4818..5009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5210..6004
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5210..6004
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5264..5283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5874..5893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6192..6211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6236..6283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttgatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggcaaacttc aagatccgcc acaacatcga ggacggcggc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagga atggagcaag tggagatcct gaggaaattc
     1381 atccagaggg tccaggccat gaagagtcct gaccacaatg gggaggacaa cttcgcccgg
     1441 gacttcatgc ggttaagaag attgtctacc aaatatagaa cagaaaagat atatcccaca
     1501 gccactggag aaaaagaaga aaatgttaaa aagaacagat acaaggacat actgccattt
     1561 gatcacagcc gagttaaatt gacattaaag actccttcac aagattcaga ctatatcaat
     1621 gcaaatttta taaagggcgt ctatgggcca aaagcatatg tagcaactca aggaccttta
     1681 gcaaatacag taatagattt ttggaggatg atatgggagt ataatgttgt gatcattgta
     1741 atggcctgcc gagaatttga gatgggaagg aaaaaatgtg agcgctattg gcctttgtat
     1801 ggagaagacc ccataacgtt tgcaccattt aaaatttctt gtgaggatga acaagcaaga
     1861 acagactact tcatcaggac actcttactt gaattcggac caggtacctg cgtggtgcac
     1921 tacaccggga tgcttgaaga tggaaagaaa tttgattcct cccgggacag aaacaagccc
     1981 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     2041 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     2101 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
     2161 ctggaaggtc ccggacaaaa tgaatctcgt aggctgtatc agtttcatta tgtgaactgg
     2221 ccagaccatg atgttccttc atcatttgat tctattctgg acatgataag cttaatgagg
     2281 aaatatcaag aacatgaaga tgttcccatt tgtattcatt gcagtgcagg ctgtggaaga
     2341 acaggtgcca tttgtgccat agattatacg tggaatttac taaaagctgg gaaaatacca
     2401 gaggaattta atgtatttaa tttaatacaa gaaatgagaa cacaaaggca ttctgcagta
     2461 caaacaaagg agcaatatga acttgttcat agagctattg cccaactgtt tgaaaaacag
     2521 ctacaactat atgaaattca tggagctcag aaaattgctg atggagtgaa tgaaattaac
     2581 actgaaaaca tgatcagctc catagagcct gaaaaacaag attctcctcc tccaaaacca
     2641 ccaaggaccc gcagttgcct tgttgaaggg gatgctaaag aagaaatact gcagccaccg
     2701 gaacctcatc cagtgccacc catcttgaca ccttctcccc cttcagcttt tccaacagtc
     2761 actactgtgt ggcaggacaa tgatagatac catccaaagc cagtgttgca tatggtttca
     2821 tcagaacaac attcagcaga cctcaacaga aactatagta aatcaacaga acttccaggg
     2881 aaaaatgaat caacaattga acagatagat aaaaaattgg aacgaaattt aagttttgag
     2941 attaagaagg tccctctcca agagggacca aaaagttttg atgggaacac acttttgaat
     3001 aggggacatg caattaaaat taaatctgct tcaccttgta tagctgataa aatctctaag
     3061 ccacaggaat taagttcaga tctaaatgtc ggtgatactt cccagaattc ttgtgtggac
     3121 tgcagtgtaa cacaatcaaa caaagtttca gttactccac cagaagaatc ccagaattca
     3181 gacacacctc caaggccaga ccgcttgcct cttgatgaga aaggacatgt aacgtggtca
     3241 tttcatggac ctgaaaatgc catacccata cctgatttat ctgaaggcaa ttcctcagat
     3301 atcaactatc aaactaggaa aactgtgagt ttaacaccaa gtcctacaac acaagttgaa
     3361 acacctgatc ttgtggatca tgataacact tcaccactct tcagaacacc cctcagtttt
     3421 actaatccac ttcactctga tgactcagac tcagatgaaa gaaactctga tggtgctgtg
     3481 acccagaata aaactaatat ttcaacagca agtgccacag tttctgctgc cactagtact
     3541 gaaagcattt ctactaggaa agtattgcca atgtccattg ctagacataa tatagcagga
     3601 acaacacatt caggtgctga aaaagatgtt gatgttagtg aagattcacc tcctccccta
     3661 cctgaaagaa ctcctgaatc gtttgtgtta gcaagtgaac ataatacacc tgtaagatcg
     3721 gaatggagtg aacttcaaag tcaggaacga tctgaacaaa aaaagtctga aggcttgata
     3781 acctctgaaa atgagaaatg tgatcatcca gcgggaggta ttcactatga aatgtgcata
     3841 gaatgtccac ctactttcag tgacaagaga gaacaaatat cagaaaatcc aacagaagcc
     3901 acagatattg gttttggtaa tcgatgtgga aaacccaaag gaccaagaga tccaccttca
     3961 gaatggacat gaggatccac cggatctaga taactgatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4681 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5221 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5281 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5341 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5401 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5461 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5521 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5581 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5641 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5701 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5761 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5821 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5881 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5941 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6001 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6061 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6121 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6181 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6241 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6421 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtatcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
