LOCUS       sequence-469415-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of FKBP-Halotag in mammalian cells. FKBP-halotag 
            localizes in the cytosol and binds to the FRB tag in the presence of
            AP21967 (A/C Heterodimerizer)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  AUTHORS   Nagashima Y, Eguchi T, Koyama-Honda I, Mizushima N
  TITLE     Optogenetic tools for inducing organelle membrane rupture.
  JOURNAL   J Biol Chem. 2025 Mar 18;301(4):108421. doi: 
            10.1016/j.jbc.2025.108421.
  PUBMED    40113041
REFERENCE   2  (bases 1 to 7133)
  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 Biol 
            Chem."; date: "2025-03-18"; volume: "301(4):108421. doi"; pages: " 
            10.1016/j.jbc.2025.108421"
COMMENT     Sequence Label: pMRX-IB-FKBP-HaloTag
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(1979..1981,1982..2302)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2303..3187
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    3215..3781
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3231..3783
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3375..3392)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3602..3621
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4459..4928
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5058..5075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5229..5817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5309..5328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5981..6772,6773..6841))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6604..6623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6842..6946)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7014..7032
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 gggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca
     2041 gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaatttg attcctcccg
     2101 ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga
     2161 agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta
     2221 tgcctatggt gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga
     2281 tgtggagctt ctaaaactgg aagaaatcgg tactggcttt ccattcgacc cccattatgt
     2341 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     2401 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     2461 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     2521 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     2581 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     2641 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     2701 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     2761 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     2821 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     2881 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     2941 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     3001 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     3061 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccaggtctga atctgctgca
     3121 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta ctctggagat
     3181 ttccggttag ctcgagagcg gccgctacgt aaattccgcc ccccccccta acgttactgg
     3241 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     3301 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     3361 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     3421 agttcctctg gaagtttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     3481 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     3541 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     3601 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     3661 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     3721 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     3781 atatggccac aaccgtcgac atggtcatga aaacatttaa catttctcaa caagatctag
     3841 aattagtaga agtagcgaca gagaagatta caatgcttta tgaggataat aaacatcatg
     3901 tgggagcggc aattcgtacg aaaacaggag aaatcatttc ggcagtacat attgaagcgt
     3961 atataggacg agtaactgtt tgtgcagaag ccattgcgat tggtagtgca gtttcgaatg
     4021 gacaaaagga ttttgacacg attgtagctg ttagacaccc ttattctgac gaagtagata
     4081 gaagtattcg agtggtaagt ccttgtggta tgtgtaggga gttgatttca gactatgcac
     4141 cagattgttt tgtgttaata gaaatgaatg gcaagttagt caaaactacg attgaagaac
     4201 tcattccact caaatatacc cgaaattaag tcgaccgccc gccccacgac ccgcagcgcc
     4261 cgaccgaaag gagcgcacga ccccatggct cgacgataaa ataaaagatt ttatttagtc
     4321 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     4381 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     4441 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     4501 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     4561 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     4621 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     4681 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     4741 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     4801 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     4861 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     4921 gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt
     4981 aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5041 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5101 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5161 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5221 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5281 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5341 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5401 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5461 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5521 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5581 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5641 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5701 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5761 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5821 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5881 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccggccgca
     5941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6001 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6061 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6121 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6181 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6241 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6301 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6361 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6421 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6481 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6541 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     6601 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6661 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6721 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6781 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6841 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6901 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6961 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7021 gtatcacgag gccctttcgt cttcactata ggagcttggc ccattgcata cgttgtatcc
     7081 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gtt
//
