LOCUS       sequence-318798-        4446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human FUS at the C-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4446)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4446)
  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"
COMMENT     Sequence Label: FUS_Halo_C_allele_400
FEATURES             Location/Qualifiers
     source          1..4446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             499..1386
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(1847..1863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1847..1863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1871..1887
                     /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(1895..1901,1902..1919,1920..1925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2078..2095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2249..2837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3008..3799,3800..3868))
                     /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     3631..3650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3869..3973)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4041..4059
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4097..4119)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4219..4238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4432..4446,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4446,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatggaccgt
       61 ggaggcttcc gagggggccg gggtggtggg gacagaggtg gctttggccc tggcaagatg
      121 gattccaggt aagactttaa atcagaataa aaaagtagag cagttgaaca gaggccatag
      181 gataacaggg ttttgttgag aaagtggttt cattttgagg gctaggtgga aagacctgag
      241 gttgtaacca gtagtggaga gggaaggaaa attaactcag ggggagtgaa tctgtagacc
      301 cacttgagat aagatactcg ctgggttagg taggaggggc agataggata tctaggcttg
      361 gagaggctgg taactcaaat ataatggata cttaattttt tttttttttt ttgcaggggt
      421 gagcacagac aggatcgcag ggagaggccg tatggtggag gaggttctgg aggcggtgga
      481 agtggtggcg gaggtagcgc cgagatcggc accggctttc ccttcgatcc tcactacgtg
      541 gaagtgctgg gcgagagaat gcactatgtg gacgtgggcc ccagagatgg aacccctgtg
      601 ctgtttctgc acggcaaccc taccagcagc tacgtgtggc ggaacatcat ccctcacgtg
      661 gcccctacac acagatgtat cgcccctgac ctgatcggca tgggcaagag cgataagccc
      721 gacctgggct acttcttcga cgaccacgtg cggttcatgg acgcctttat tgaggccctc
      781 ggcctggaag aggtggtgct ggttattcac gattggggca gcgccctggg attccactgg
      841 gccaagagaa accccgagag agtgaaggga atcgccttca tggaattcat caggcccatt
      901 cctacctggg acgagtggcc tgagttcgcc agagagacat tccaggcctt cagaaccacc
      961 gacgtgggca gaaagctgat catcgaccag aacgtgttca tcgagggcac cctgcctatg
     1021 ggagtcgtca gaccactgac cgaggtggaa atggaccact acagagagcc ctttctgaac
     1081 cccgtggacc gggaacctct ttggagattc cccaacgagc tgcccattgc cggcgagcct
     1141 gctaatattg tggccctggt ggaagagtac atggactggc tgcatcagag ccccgtgcct
     1201 aagctgctgt tttggggaac acccggcgtg ctgattcctc ctgctgaagc tgccagactg
     1261 gccaagagcc tgcctaattg caaggccgtg gatatcggcc ctggcctgaa tctgctgcaa
     1321 gaggacaacc ccgatctgat cggctccgag attgccagat ggctgagcac cctggaaatc
     1381 agcggataat tagcctggct ccccaggttc tggaacagct ttttgtcctg tacccagtgt
     1441 taccctcgtt attttgtaac cttccaattc ctgatcaccc aagggttttt ttgtgtcgga
     1501 ctatgtaatt gtaactatac ctctggttcc cattaaaagt gaccatttta gttaaatttt
     1561 gttcctcttc ccccttttca ctttcctgga agatcgatgt cccgatcagg aaggtagaga
     1621 gttttcctgt tcagattacc ctgcccagca ggaactggaa tacagtgttc ggggagaagg
     1681 ccaaatgata tccttgagag cagagattaa acttttctgt catggggaaa gttggtgtat
     1741 aaatgagaaa tgaagaacat gggatgtcat gagtgttggc ctaaatttga tcggatcccg
     1801 ggcccgtcga ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc
     1861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3121 ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag
     3181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3961 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4021 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4081 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4141 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4201 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     4261 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     4321 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4381 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4441 gacgtt
//
