LOCUS       sequence-369575-        7003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin regulated PTP1B with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7003)
  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 7003)
  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: flag-RapR-PTP1B
FEATURES             Location/Qualifiers
     source          1..7003
                     /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      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2556..2574)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2557..2574)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2594..2611)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2600..2824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2870..3298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2957..2976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3167..3188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3307..3327)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3312..3641
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3492..3627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3554..3573
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3708..4502
                     /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(3762..3781)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4372..4391
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4676..4797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4713..4732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4767..4786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4846..4862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4846..4862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4859..4881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4870..4886
                     /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(4894..4900,4901..4918,4919..4924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4939..4960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5077..5094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6007..6798,6799..6867))
                     /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     6630..6649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6868..6972)
                     /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 gcttgccacc
      901 atggattaca aggatgacga cgataagatc gaattcatgg agatggaaaa ggagttcgag
      961 cagatcgaca agtccgggag ctgggcggcc atttaccagg atatccgaca tgaagccagt
     1021 gacttcccat gtagagtggc caagcttcct aagaacaaaa accgaaatag gtacagagac
     1081 gtcagtccct ttgaccatag tcggattaaa ctacatcaag aagataatga ctatatcaac
     1141 gctagtttga taaaaatgga agaagcccaa aggagttaca ttcttaccca gggccctttg
     1201 cctaacacat gcggtcactt ttgggagatg gtgtgggagc agaaaagcag gggtgtcgtc
     1261 atgctcaaca gagtgatgga gaaaggttcg ttaaaatgcg cacaatactg gccacaaaaa
     1321 gaagaaaaag agatgatctt tgaagacaca aatttgaaat taacattgat ctctgaagat
     1381 atcaagtcat attatacagt gcgacagcta gaattggaaa accttggacc aggtacctgc
     1441 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     1501 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     1561 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     1621 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     1681 cttctaaaac tggaaggtcc cggaacccaa gaaactcgag agatcttaca tttccactat
     1741 accacatggc ctgactttgg agtccctgaa tcaccagcct cattcttgaa ctttcttttc
     1801 aaagtccgag agtcagggtc actcagcccg gagcacgggc ccgttgtggt gcactgcagt
     1861 gcaggcatcg gcaggtctgg aaccttctgt ctggctgata cctgcctctt gctgatggac
     1921 aagaggaaag acccttcttc cgttgatatc aagaaagtgc tgttagaaat gaggaagttt
     1981 cggatggggc tgatccagac agccgaccag ctgcgcttct cctacctggc tgtgatcgaa
     2041 ggtgccaaat tcatcatggg ggactcttcc gtgcaggatc agtggaagga gctttcccac
     2101 gaggacctgg agcccccacc cgagcatatc cccccacctc cccggccacc caaacgaatc
     2161 ctggagccac acaatgggaa atgcagggag ttcttcccaa atcaccagtg ggtgaaggaa
     2221 gagacccagg aggataaaga ctgccccatc aaggaagaaa aaggaagccc cttaaatgcc
     2281 gcaccctacg gcatcgaaag catgagtcaa gacactgaag ttagaagtcg ggtcgtgggg
     2341 ggaagtcttc gaggtgccca ggctgcctcc ccagccaaag gggagccgtc actgcccgag
     2401 aaggacgagg accatgcact gagttactgg aagcccttcc tggtcaacat gtgcgtggct
     2461 acggtcctca cggccggcgc ttacctctgc tacaggttcc tgttcaacag caacacatag
     2521 gcggccgcgt cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     2581 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2641 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2701 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2761 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2821 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2881 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2941 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3001 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3061 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3121 tagacggttt ttcgcccttt gacgttggag tccacgttcg ttaatagtgg actcttgttc
     3181 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3241 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3301 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3361 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3481 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3541 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3601 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3661 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3721 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3781 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3841 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3901 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3961 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4021 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4081 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4141 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4201 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4261 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4321 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4381 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4441 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4501 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4561 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4621 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4681 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4741 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4801 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4861 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4921 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4981 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
