LOCUS       sequence-349366-        4374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human CHCHD10 at the 
            N-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4374)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4374)
  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: CHCHD10_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(431..447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(431..447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(444..466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    455..471
                     /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(479..485,486..503,504..509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    524..545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(662..679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(833..1421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(913..932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1592..2383,2384..2452))
                     /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     2215..2234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2453..2557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2625..2643
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2681..2703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2803..2822
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3016..3038
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3030..3047
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3031..3047
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3448..3457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3454..4344
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
ORIGIN
        1 ggcggaggta gccctcgggg aagccgcagc gcggcctccc ggccagccag gtgtgagggg
       61 gacaagggcc ctgtggggag ggggttaggg aagtgtgggc actcccctaa gggcgtccgc
      121 agagtgacct tggctgctgc tgcgccatct tggcgggggg aggggtgggg gtgcagcacc
      181 cacgtgtgga gacgtgggtc gtccatctct ccgggggtcg tttccaggag ctgcagccct
      241 ccagcctggg gtctggccag gattaaccct gcttcctccc acccccgcag ccgcccagcc
      301 gcgccctctg cccacccgcc cgcgcaccca ccgccctcgg cagccgcccc agcccccgcc
      361 ccttcgggcc agccggggct catggctcag atggcgacca cggccgcagg ggaagcttgg
      421 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      481 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1261 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac
     1741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2581 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2641 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2701 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2761 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     2821 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     2881 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     2941 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3001 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcccccgtc
     3061 tcctctcccc tccagggctc gcaacaaccc ccaactccgc tccctgccct ctgcggtggt
     3121 cctaacagaa ggcactgtcg ccctccttaa gccctcggcg ctttgcgaca ctggcccctg
     3181 tccccagaac ccgcacagcc tgccaggcgc agggagggag cagggtccca tttccagccg
     3241 agacccggag ctgctggaag ggagatgtcc ggaggacggg agaaggggga tagggttggg
     3301 ggcagtgccg cctcccgctg ggggcgtgtt cctgaggccc cgcccggggc cccgcctttt
     3361 ttgcgccggc tgtgacaagc gctgcggcat ttgtccccgc gacagcaccg ctgccgccgt
     3421 ctctaaggtc gcccgggtcc caccgccgcc accatggccg agatcggcac cggctttccc
     3481 ttcgatcctc actacgtgga agtgctgggc gagagaatgc actatgtgga cgtgggcccc
     3541 agagatggaa cccctgtgct gtttctgcac ggcaacccta ccagcagcta cgtgtggcgg
     3601 aacatcatcc ctcacgtggc ccctacacac agatgtatcg cccctgacct gatcggcatg
     3661 ggcaagagcg ataagcccga cctgggctac ttcttcgacg accacgtgcg gttcatggac
     3721 gcctttattg aggccctcgg cctggaagag gtggtgctgg ttattcacga ttggggcagc
     3781 gccctgggat tccactgggc caagagaaac cccgagagag tgaagggaat cgccttcatg
     3841 gaattcatca ggcccattcc tacctgggac gagtggcctg agttcgccag agagacattc
     3901 caggccttca gaaccaccga cgtgggcaga aagctgatca tcgaccagaa cgtgttcatc
     3961 gagggcaccc tgcctatggg agtcgtcaga ccactgaccg aggtggaaat ggaccactac
     4021 agagagccct ttctgaaccc cgtggaccgg gaacctcttt ggagattccc caacgagctg
     4081 cccattgccg gcgagcctgc taatattgtg gccctggtgg aagagtacat ggactggctg
     4141 catcagagcc ccgtgcctaa gctgctgttt tggggaacac ccggcgtgct gattcctcct
     4201 gctgaagctg ccagactggc caagagcctg cctaattgca aggccgtgga tatcggccct
     4261 ggcctgaatc tgctgcaaga ggacaacccc gatctgatcg gctccgagat tgccagatgg
     4321 ctgagcaccc tggaaatcag cggaggagga ggttctggag gcggtggaag tggt
//
