LOCUS       sequence-318796-        4437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human FUS at the N-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4437)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4437)
  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_N_allele_400
FEATURES             Location/Qualifiers
     source          1..4437
                     /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             448..1338
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1851..1873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1862..1878
                     /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(1886..1892,1893..1910,1911..1916))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1931..1952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2069..2086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2240..2828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2320..2339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2999..3790,3791..3859))
                     /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     3622..3641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3860..3964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4032..4050
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4088..4110)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4210..4229
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4423..4437,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4437,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatccagccg
       61 ccgggaccgc cccctcgccc cgcccccagc gggcactcag gccgtaccac tgtgccttca
      121 tgggggtgga gatagatcgt gggctagtcc tgccgaggag agaggggttc ttcctcaaaa
      181 aatatgatta tgtatagtat tcgcatgatt ctagttaact tgtttccctt ctgcctgctc
      241 ggaccctcta cctgccctac gaagggggcg gagtgcgttc ctgcctcccc ctgctcttcc
      301 gcgtttggtg cgcgcctgcg cggtgcgtag gcggcggagc gtacttaagc ttcgacgcag
      361 gaggcggggc tgctcagtcc tccaggcgtc ggtactcagc ggtgttggaa cttcgttgct
      421 tgcttgcctg tgcgcgcgtg cgcggacatg gccgagatcg gcaccggctt tcccttcgat
      481 cctcactacg tggaagtgct gggcgagaga atgcactatg tggacgtggg ccccagagat
      541 ggaacccctg tgctgtttct gcacggcaac cctaccagca gctacgtgtg gcggaacatc
      601 atccctcacg tggcccctac acacagatgt atcgcccctg acctgatcgg catgggcaag
      661 agcgataagc ccgacctggg ctacttcttc gacgaccacg tgcggttcat ggacgccttt
      721 attgaggccc tcggcctgga agaggtggtg ctggttattc acgattgggg cagcgccctg
      781 ggattccact gggccaagag aaaccccgag agagtgaagg gaatcgcctt catggaattc
      841 atcaggccca ttcctacctg ggacgagtgg cctgagttcg ccagagagac attccaggcc
      901 ttcagaacca ccgacgtggg cagaaagctg atcatcgacc agaacgtgtt catcgagggc
      961 accctgccta tgggagtcgt cagaccactg accgaggtgg aaatggacca ctacagagag
     1021 ccctttctga accccgtgga ccgggaacct ctttggagat tccccaacga gctgcccatt
     1081 gccggcgagc ctgctaatat tgtggccctg gtggaagagt acatggactg gctgcatcag
     1141 agccccgtgc ctaagctgct gttttgggga acacccggcg tgctgattcc tcctgctgaa
     1201 gctgccagac tggccaagag cctgcctaat tgcaaggccg tggatatcgg ccctggcctg
     1261 aatctgctgc aagaggacaa ccccgatctg atcggctccg agattgccag atggctgagc
     1321 accctggaaa tcagcggagg aggaggttct ggaggcggtg gaagtggtgg cggaggtagc
     1381 tcaaacggta ggtaagggcg cgaggcgacg gcggcggcgc acccggccga ggcctcccag
     1441 ctgggctttt cgttttcagt gggaccgggg cggcgatccc gtgtgggatt ttttggcgcc
     1501 cctgtggcgg gaagccgcgg agaagagtaa ctggaggagg ctggtgtcgc cattttgttt
     1561 cgctcctctg gccctcgcgc gcggggcggg aagtcttttc tttgcagtcc gtttgcttgg
     1621 ggtgggcgtt gggagggacg cttcttaggg gtttgaagcg tcaggtgagg gtggaaaacg
     1681 cccattctcc gtggcctcgc ctcccccaac tcccggcccc gcgctcgagc ccgctttgtc
     1741 gcagtgctgc atccgggcac tcgcggcgcg cacgcgctct atcggatccc gggcccgtcg
     1801 actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3121 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     3181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3961 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4021 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4081 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4141 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4201 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     4261 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     4321 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     4381 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
