LOCUS       sequence-451304-        5472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of LgBiT-MYPT1(1--314).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5472)
  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 5472)
  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-LgBiT-MYPT1(1-314)(KAKA)-Flag
FEATURES             Location/Qualifiers
     source          1..5472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    115..236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(152..171)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     206..225
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(243..698)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(380..401)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     592..611
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        725..829
                     /gene="bla"
                     /label=AmpR promoter
     promoter        831..1188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        831..1022
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(854..874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1039..1174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1223..2017
                     /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             1223..2017
                     /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(1277..1296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1887..1906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2205..2224)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2249..2296
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2625..3213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3114..3133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3388..3691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3692..3895
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3846..3866
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5446..5469
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
       61 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      121 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      181 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
      241 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      301 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      361 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      421 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      481 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      541 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      601 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      661 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
      721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
      841 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     1201 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     1261 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     1321 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     1381 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     1441 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     1501 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     1561 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1621 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     1681 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     1741 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     1801 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1861 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     1921 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1981 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     2041 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     2101 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     2161 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     2221 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     2281 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2341 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2401 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2461 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3301 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     3361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3421 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3481 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3541 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3601 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3661 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3721 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3781 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3841 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     3901 agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttcgaatt
     3961 accatggtct tcacactcga agatttcgtt ggggactggg aacagacagc cgcctacaac
     4021 ctggaccaag tccttgaaca gggaggtgtg tccagtttgc tgcagaatct cgccgtgtcc
     4081 gtaactccga tccaaaggat tgtccggagc ggtgaaaatg ccctgaagat cgacatccat
     4141 gtcatcatcc cgtatgaagg tctgagcgcc gaccaaatgg cccagatcga agaggtgttt
     4201 aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcccta tggcacactg
     4261 gtaatcgacg gggttacgcc gaacatgctg aactatttcg gacggccgta tgaaggcatc
     4321 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt
     4381 atcgacgagc gcctgatcac ccccgacggc tccatgctgt tccgagtaac catcaacagt
     4441 gggagttccg gtggtggcgg gagcggaggt ggaggctcga gcggtatggc ggatgcggac
     4501 gcgaagcaga agcggaacga gcagctgaaa cgctggatcg gctccgagac ggacctcgag
     4561 cctccggtgg tgaagcgcca gaagaccaag gcgaaggccg acgatggcgc cgtcttcctg
     4621 gctgcttgct ccagcggcga cacggacgag gtcctcaagc tgctgcaccg cggcgccgac
     4681 atcaattacg ccaatgtgga cggactcact gccctgcacc aggcttgcat tgatgacaat
     4741 gttgatatgg tgaagtttct ggtagaaaat ggagcaaata ttaatcaacc tgataatgaa
     4801 ggctggatac cactacatgc agcagcttcc tgtggatatc ttgatattgc agagtttttg
     4861 attggtcaag gagcacatgt aggggctgtc aacagtgaag gagatacacc tttagatatt
     4921 gcggaggagg aggcaatgga agagctactt caaaatgaag ttaatcggca aggggttgat
     4981 atagaagcag ctcgaaagga agaagaacgg atcatgctta gagatgccag gcagtggcta
     5041 aatagtggtc atataaatga tgtccggcat gcaaaatctg gaggtacagc acttcacgtt
     5101 gcagctgcta aaggctatac ggaagtttta aaacttttaa tacaggcagg ctatgatgtt
     5161 aatattaaag actatgatgg ctggacacct cttcatgctg cagctcattg gggtaaagaa
     5221 gaagcatgtc gaattttagt ggacaatctg tgtgatatgg agatggtcaa caaagtgggc
     5281 caaacagcct ttgatgtagc agatgaagac attttaggat atttagaaga gttgcaaaag
     5341 aaacaaaatc tgctccatag tgaaaaacgg gacaagaaat ctccactaat tgaatcaaca
     5401 gcaaatatgg acaataatca gtcacagaag ggcagcacca gcggcgacta caaagacgat
     5461 gacgacaagt aa
//
