LOCUS       sequence-349361-        4436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human LRRK2 at the N-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4436)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4436)
  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: LRRK2_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2048..2070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2170..2189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2383..2405
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2397..2414
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2398..2414
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2844..2853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2850..3740
                     /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(4234..4250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4234..4250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4247..4269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4258..4274
                     /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(4282..4288,4289..4306,4307..4312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4327..4348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     2221 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     2281 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     2341 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     2401 aaacgacggc cagtgaattc gagctcggta cctcgcgaat gcatctagag gcgtccgccc
     2461 ggggtccggt ctaggcgtgc gtgggggcca cggtcacggt catcccagcc aggcccggct
     2521 ccagcagccc cacggccgcc gccagagttc tgcgcggccc gtcgcctcgg cggagcctct
     2581 ggcaggcccc tgagctcgtt tttggggcct gagtggggga ggaggaagcc gagcaggagg
     2641 gctccggaga gggagggcaa cgcggggcgg ggagctgcct ccttcctcat aaacaggcgg
     2701 gcgtgggcgc cgatggggcc cgcggggagc gctggctgcg ggcggtgagc tgagctcgcc
     2761 cccggggagc tgtggccggc gcccctgccg gttccctgag cagcggacgt tcatgctggg
     2821 agggcggcgg gttggaagca ggtgccacca tggctgagat cggcaccggc tttcccttcg
     2881 atcctcacta cgtggaagtg ctgggcgaga gaatgcacta tgtggacgtg ggccccagag
     2941 atggaacccc tgtgctgttt ctgcacggca accctaccag cagctacgtg tggcggaaca
     3001 tcatccctca cgtggcccct acacacagat gtatcgcccc tgacctgatc ggcatgggca
     3061 agagcgataa gcccgacctg ggctacttct tcgacgacca cgtgcggttc atggacgcct
     3121 ttattgaggc cctcggcctg gaagaggtgg tgctggttat tcacgattgg ggcagcgccc
     3181 tgggattcca ctgggccaag agaaaccccg agagagtgaa gggaatcgcc ttcatggaat
     3241 tcatcaggcc cattcctacc tgggacgagt ggcctgagtt cgccagagag acattccagg
     3301 ccttcagaac caccgacgtg ggcagaaagc tgatcatcga ccagaacgtg ttcatcgagg
     3361 gcaccctgcc tatgggagtc gtcagaccac tgaccgaggt ggaaatggac cactacagag
     3421 agccctttct gaaccccgtg gaccgggaac ctctttggag attccccaac gagctgccca
     3481 ttgccggcga gcctgctaat attgtggccc tggtggaaga gtacatggac tggctgcatc
     3541 agagccccgt gcctaagctg ctgttttggg gaacacccgg cgtgctgatt cctcctgctg
     3601 aagctgccag actggccaag agcctgccta attgcaaggc cgtggatatc ggccctggcc
     3661 tgaatctgct gcaagaggac aaccccgatc tgatcggctc cgagattgcc agatggctga
     3721 gcaccctgga aatcagcgga ggaggaggtt ctggaggcgg tggaagtggt ggcggaggta
     3781 gcgctagtgg cagctgtcag gggtgcgaag aggacgagga aactctgaag aagttgatag
     3841 tcaggctgaa caatgtccag gaaggaaaac agatagaaac gctggtccaa atcctggagg
     3901 atctgctggt gttcacgtac tccgagcacg gtaatcactt gaaaataaac tgtgctttta
     3961 tttttgcaaa ctttctcccc ctccttacat ttgcaaattt tgtcctcctc cccttgaccc
     4021 tgctcaaacc cggactctta aggagccgca aactcccata tcctttcctt agggcagaaa
     4081 gcagctgaga atttcaggaa ggtcttcacc tttttgactt ttctccccgt ttcagactaa
     4141 aaaggaaagg gggtgctgtg gattgtgact ttgcttcttg gatcccgggc ccgtcgactg
     4201 cagaggcctg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4261 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4321 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4381 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcg
//
