LOCUS       sequence-349342-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human TMEM106B at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4377)
  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: TMEM106B_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(158..746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(238..257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(917..1708,1709..1777))
                     /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     1540..1559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1778..1882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1950..1968
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2006..2028)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2128..2147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2341..2363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2355..2372
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2356..2372
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2824..3711
                     /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(4133..4149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4133..4149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4146..4168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4157..4173
                     /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(4181..4187,4188..4205,4206..4211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4226..4247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4364..4377,1..4))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
       61 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1021 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
     1081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1921 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     1981 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2041 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2101 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2161 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     2221 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2281 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2341 cccagtcacg acgttgtaaa acgacggcca gtgaattcga acttttatat ttttaacaga
     2401 agataattgt tttaaaatat tacacttatt atgtgtaatt atgtctacag ggctcactca
     2461 gctatccatt tttgttgtct gttggggaaa tactccttaa gaggattgtg tgcacaatat
     2521 taagttatca ttaatcaaat attctcttct gggagataat ttttatgtgt taaagtagtc
     2581 tcactatgga aaaacttcta atataactat taaatgtctc tcctcactta cattattttt
     2641 agagttactg tgacaacaac atactttggc cactctgaac agatatccca ggagaggtat
     2701 cagtatgtcg actgtggaag aaacacaact tatcagttgg ggcagtctga atatttaaat
     2761 gtacttcagc cacaacaggg tggaggaggt tctggaggcg gtggaagtgg tggcggaggt
     2821 agcgccgaga tcggcaccgg ctttcccttc gatcctcact acgtggaagt gctgggcgag
     2881 agaatgcact atgtggacgt gggccccaga gatggaaccc ctgtgctgtt tctgcacggc
     2941 aaccctacca gcagctacgt gtggcggaac atcatccctc acgtggcccc tacacacaga
     3001 tgtatcgccc ctgacctgat cggcatgggc aagagcgata agcccgacct gggctacttc
     3061 ttcgacgacc acgtgcggtt catggacgcc tttattgagg ccctcggcct ggaagaggtg
     3121 gtgctggtta ttcacgattg gggcagcgcc ctgggattcc actgggccaa gagaaacccc
     3181 gagagagtga agggaatcgc cttcatggaa ttcatcaggc ccattcctac ctgggacgag
     3241 tggcctgagt tcgccagaga gacattccag gccttcagaa ccaccgacgt gggcagaaag
     3301 ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ctatgggagt cgtcagacca
     3361 ctgaccgagg tggaaatgga ccactacaga gagccctttc tgaaccccgt ggaccgggaa
     3421 cctctttgga gattccccaa cgagctgccc attgccggcg agcctgctaa tattgtggcc
     3481 ctggtggaag agtacatgga ctggctgcat cagagccccg tgcctaagct gctgttttgg
     3541 ggaacacccg gcgtgctgat tcctcctgct gaagctgcca gactggccaa gagcctgcct
     3601 aattgcaagg ccgtggatat cggccctggc ctgaatctgc tgcaagagga caaccccgat
     3661 ctgatcggct ccgagattgc cagatggctg agcaccctgg aaatcagcgg ataaaaactg
     3721 gaagagatgg atttaaagaa gaaatatcta ttgatatttc ctatactctc aatgaagagg
     3781 tatttcctaa taggagacct taaattgaac aaacctaaag tttacacttc taagagtaca
     3841 gttaaaagta tgtggacctg cagttcttgt aactctccac tctgtgttaa tgatatattt
     3901 gtactaggat cttttacttg aatctaaatt tactggttga tttccttctc cagcctatcc
     3961 cctacaggga aaagctgata cttcccctat agtacaataa ataattattt aaaagtcata
     4021 gctccagtca ctactgaaaa cataattttg gtgataaaca taatttgaga aacttaattt
     4081 ctgaatgttt ttatagaaaa ttactgaaag tctaaagctt ggcgtaatca tggtcatagc
     4141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgact
//
