LOCUS       sequence-260188-        5333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Halo Tag, Mitochondrial outer membrane localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5333)
  AUTHORS   Wang C, Taki M, Sato Y, Tamura Y, Yaginuma H, Okada Y, Yamaguchi S
  TITLE     A photostable fluorescent marker for the superresolution live 
            imaging of the dynamic structure of the mitochondrial cristae.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Aug 6;116(32):15817-15822. doi: 
            10.1073/pnas.1905924116. Epub 2019 Jul 23.
  PUBMED    31337683
REFERENCE   2  (bases 1 to 5333)
  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: 
            "10.1073/pnas.1905924116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-08-6- 6"; volume: "116"; issue: "32"; pages: 
            "15817-15822"
COMMENT     Sequence Label: pN1-TOMM20-Halo
FEATURES             Location/Qualifiers
     source          1..5333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      575..584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1052..1936
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    2061..2182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2098..2117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2152..2171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2189..2644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2538..2557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2671..2775
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2777..3134
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2777..2968
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2800..2820)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2985..3120
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3169..3963
                     /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             3169..3963
                     /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(3223..3242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3833..3852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4151..4170)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4195..4242
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4571..5159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5060..5079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgccacc atggtgggtc ggaacagcgc
      601 catcgccgcc ggtgtatgcg gggccctttt cattgggtac tgcatctact tcgaccgcaa
      661 aagacgaagt gaccccaact tcaagaacag gcttcgagaa cgaagaaaga aacagaagct
      721 tgccaaggag agagctgggc tttccaagtt acctgacctt aaagatgctg aagctgttca
      781 gaagttcttc cttgaagaaa tacagcttgg tgaagagtta ctagctcaag gtgaatatga
      841 gaagggcgta gaccatctga caaatgcaat tgctgtgtgt ggacagccac agcagttact
      901 gcaggtctta cagcaaactc ttccaccacc agtgttccag atgcttctga ctaagctccc
      961 aacaattagt cagagaattg taagtgctca gagcttggct gaagatgatg tggaaggcgg
     1021 tagcggggat ccaccggtct cgaccggcag cgaaatcggt actggctttc cattcgaccc
     1081 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
     1141 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
     1201 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
     1261 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
     1321 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     1381 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     1441 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     1501 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     1561 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     1621 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     1681 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     1741 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     1801 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     1861 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac
     1921 tctggagatt tccggttaag cggccgcgac tctagatcat aatcagccat accacatttg
     1981 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2041 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2101 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2161 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     2221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2281 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2401 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2641 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2701 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2761 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2821 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2881 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2941 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3001 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3061 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3121 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3961 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4141 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4201 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4261 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4321 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4381 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     4441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     5281 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccg
//
