LOCUS       sequence-451306-        5121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Flag-SmBiT-PP1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5121)
  AUTHORS   Claes Z, Bollen M
  TITLE     A split-luciferase lysate-based approach to identify small-molecule 
            modulators of phosphatase subunit interactions.
  JOURNAL   Cell Chem Biol. 2023 Dec 21;30(12):1666-1679.e6. doi: 
            10.1016/j.chembiol.2023.07.018. Epub 2023 Aug 24.
  PUBMED    37625414
REFERENCE   2  (bases 1 to 5121)
  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: 
            "10.1016/j.chembiol.2023.07.018"; journalName: "Cell Chem Biol"; 
            date: "2023-12-21- 21"; volume: "30"; issue: "12"; pages: 
            "1666-1679.e6"
COMMENT     Sequence Label: pEGFP-C1-Flag-SmBiT-PP1b
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1105..1226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1142..1161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1196..1215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1233..1688)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1370..1391)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1582..1601
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1715..1819
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1821..2178
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1821..2012
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1844..1864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2029..2164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2213..3007
                     /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             2213..3007
                     /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(2267..2286)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2877..2896
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3195..3214)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3239..3286
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3615..4203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4104..4123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4378..4681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4682..4885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4836..4856
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4957..4980
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 atggcggacg gggagctgaa cgtggacagc ctcatcaccc gcctgctgga ggtacgagga
       61 tgtcgtccag gaaagattgt gcagatgact gaagcggaag tgcggggcct ttgtatcaag
      121 tctcgggaga tcttcctcag ccagcctatt ctcttggaat tggaagcacc actgaaaatt
      181 tgtggagata ttcatggaca gtatacagat ttactgagat tatttgagta tggaggtttc
      241 ccaccagaag ccaactatct tttcttagga gattatgtgg atagaggaaa acagtctttg
      301 gaaaccattt gtttgctatt ggcttacaaa atcaaatatc cagagaactt ctttctctta
      361 agaggaaacc atgagtgtgc tagcatcaat cgcatttatg gattctatga tgaatgcaag
      421 cgaaggttta atattaaatt gtggaagacc ttcactgact gctttaactg tctgcctata
      481 gctgccattg tggatgagaa gatcttctgt tgtcatggag gactgtcacc agacctacaa
      541 tctatggagc agattcggag aataatgagg cccacagacg tccctgatac aggtttgctc
      601 tgtgatttac tgtggtctga cccagacaag gacgtgcaag gctggggaga aaatgaccgt
      661 ggtgtttcct tcacttttgg cgctgatgta gtcagtaaat ttctgaatcg tcatgattta
      721 gacctgattt gtcgagctca tcaggtggtg gaagatggat atgaattttt tgctaagcga
      781 caattggtaa ccctattttc agccccaaat tactgtggcg agtttgataa tgctggtgga
      841 atgatgagtg tggatgagac tttgatgtgt tcatttcaga tattgaaacc atctgaaaag
      901 aaagctaaat accagtatgg tggactgaat tctggacgcc ctgtcactcc acctcgaaca
      961 gctaatccac cgaagaaaag gccgcggctt cgactctaga tcataatcag ccataccaca
     1021 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     1081 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     1141 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1201 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     1261 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1321 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1381 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1441 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1501 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1561 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1621 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1681 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1741 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1801 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     1861 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1921 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1981 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2041 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2101 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2161 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     2221 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2281 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2341 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     2401 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2461 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2521 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2581 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2641 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2701 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     2761 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2821 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2881 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2941 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3001 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3061 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3121 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     3181 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3241 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     3301 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3361 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3421 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     3481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3601 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3781 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4201 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4261 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     4321 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4381 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4441 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4501 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4561 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4621 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4681 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4741 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4801 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4861 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     4921 gactcagatc tcgagctcaa gcttcgaatt accatggact acaaagacga tgacgacaag
     4981 ggaggatccg gcggcgtctt cacactcgaa gatttcggcg tgaccggcta ccggctgttc
     5041 gaggagattc tggtaaccat caacagtggg agttccggtg gtggcgggag cggaggtgga
     5101 ggctcgagcg gtatggcgga t
//
