LOCUS       sequence-213892-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hFis with selflabeling tag, e.g. for fluorescence microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  AUTHORS   Appelhans T, Richter CP, Wilkens V, Hess ST, Piehler J, Busch KB
  TITLE     Nanoscale organization of mitochondrial microcompartments revealed 
            by combining tracking and localization microscopy.
  JOURNAL   Nano Lett. 2012 Feb 8;12(2):610-6. doi: 10.1021/nl203343a. Epub 2012
            Jan 13.
  PUBMED    22201267
REFERENCE   2  (bases 1 to 6584)
  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: "10.1021/nl203343a";
            journalName: "Nano Lett"; date: "2012-02-8- 8"; volume: "12"; issue:
            "2"; pages: "610-6"
COMMENT     Sequence Label: pSEMS-Halo7Tag-hFis
FEATURES             Location/Qualifiers
     source          1..6584
                     /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"
     CDS             936..1820
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     intron          2375..2604
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2587..2606
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2662..3235
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2829..2846)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3056..3075
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3271..4074
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3334..3353)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3944..3963
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4204..4325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4241..4260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4427..4443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4440..4462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4451..4467
                     /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(4475..4481,4482..4499,4500..4505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4520..4541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4658..4675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4829..5417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4909..4928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5588..6379,6380..6448))
                     /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     6211..6230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6449..6553)
                     /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 gcttggtacc
      901 gagctcggat cgatatcgaa ttccatatgg gatccgaaat cggtactggc tttccattcg
      961 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1021 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1081 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1141 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1201 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1261 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1321 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1381 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1441 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1501 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1561 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1621 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1681 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1741 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     1801 ctactctgga gatttccggt cctgcaggta taggcgcgcc aggatccatg gaggccgtgc
     1861 tgaacgagct ggagtctgtg gaggacctgc tgaagtttga aaagaaattt cagtctgaga
     1921 aggcagcagg ctcggtgtcc aagagcacgc agtttgagta cgcctggtgc ctggtgcgga
     1981 gcaagtacaa tgatgacatc cgtaaaggca tcgtgctgct cgaggagctg ctgcccaaag
     2041 ggagcaagga ggaacagcgg gattacgtct tctacctggc cgtggggaac taccggctca
     2101 aggaatacga gaaggcctta aagtacgtcc gcgggttgct gcagacagag ccccagaaca
     2161 accaggccaa ggaactggag cggctcattg acaaggccat gaagaaagat ggactcgtgg
     2221 gcatggccat cgtgggaggc atggccctgg gtgtggcggg actggccgga ctcatcggac
     2281 ttgctgtgtc caagtccaaa tcctgagcgg ccgcatagat aactgatcca gtgtgctgga
     2341 attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc aaaagcgggc
     2401 atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg
     2461 cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg
     2521 ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca
     2581 tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag gtcgagcatg
     2641 catctagggc ggccaattcc gcccctctcc cccccccccc cctaacgtta ctggccgaag
     2701 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     2761 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     2821 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     2881 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2941 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     3001 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     3061 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     3121 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     3181 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg
     3241 ccacaacccg ggataattcc tgcagccaat atgggatcgg ccattgaaca agatggattg
     3301 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3361 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3421 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3481 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3541 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3601 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3661 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3721 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3781 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc
     3841 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3901 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3961 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4021 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaggggat
     4081 caattctcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     4141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4321 tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4381 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4441 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4501 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4561 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4621 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4681 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4741 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4801 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4861 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4921 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4981 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5041 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5101 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5161 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5221 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5281 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5341 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5581 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5641 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5701 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5761 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5821 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5881 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5941 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6001 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6061 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6121 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6181 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6241 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6301 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6361 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6421 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
