LOCUS       sequence-349320-        4453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human COQ2 at the N-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4453)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4453)
  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: COQ2_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(413..429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(413..429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(426..448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    437..453
                     /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(461..467,468..485,486..491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    506..527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(644..661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(815..1403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(895..914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1574..2365,2366..2434))
                     /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     2197..2216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2435..2539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2607..2625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2663..2685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2785..2804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2998..3020
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3012..3029
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3013..3029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3471..4361
                     /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 ggcgtggctg ccgggctggc ggggccgctc cttcgccctg gcgcgtgcgg caggcgcgcc
       61 ccacggtggt gacttgcagc cccccgcctg tcccgagccg cgcgggcgcc agctcagttt
      121 gtccgcggcg gcggtggtgg actctgcgcc ccgccccctg cagccgtact tgcgcctcat
      181 gcggttggac aagcccattg gtgagtgcgg gcgggcgggc agcccgggaa tttgcaagta
      241 gcagccgccg agtcggctcc gcggagctgt ccgcggcggc cggccggggc gtgatggaaa
      301 tgagaacctg aaagcttggg cttggctgcc gggtgccgtg cgccctgggg ccaatatcgg
      361 atcccgggcc cgtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc
      421 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      481 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1681 taccatctgg ccccagtgct gcaatgatac cgcgagatcc acgctcaccg gctccagatt
     1741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2581 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2641 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2701 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2761 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2821 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2881 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2941 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3001 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacctcg cgaatgcatc
     3061 tagatccaat ataaagccaa cttcctccgc tcgactcttt ggaacactta ttctatttta
     3121 cagaataagg tgttgcccga ttctattttt tgtttgtttc ctcagagacc ccgttctgag
     3181 tgttgcccga taatggaacc acaaagtcaa tcaagatcga gttgtaaaaa attacaacta
     3241 aatttattgt aatttttttg ccttttgcca atagaaatcc ctttttcatt aagtattcct
     3301 ccctagagta agcgaccacg atgaccccaa tttcacaagt aaggatgagg aaaggttctg
     3361 cccacaccgc cgcccagcct ggcagactag gattgcatcc tgcgggtgcc actgcgcatg
     3421 cctgccgggg aatgacgtca atccgagctc gtcccggcct caccagcgcc atggccgaga
     3481 tcggcaccgg ctttcccttc gatcctcact acgtggaagt gctgggcgag agaatgcact
     3541 atgtggacgt gggccccaga gatggaaccc ctgtgctgtt tctgcacggc aaccctacca
     3601 gcagctacgt gtggcggaac atcatccctc acgtggcccc tacacacaga tgtatcgccc
     3661 ctgacctgat cggcatgggc aagagcgata agcccgacct gggctacttc ttcgacgacc
     3721 acgtgcggtt catggacgcc tttattgagg ccctcggcct ggaagaggtg gtgctggtta
     3781 ttcacgattg gggcagcgcc ctgggattcc actgggccaa gagaaacccc gagagagtga
     3841 agggaatcgc cttcatggaa ttcatcaggc ccattcctac ctgggacgag tggcctgagt
     3901 tcgccagaga gacattccag gccttcagaa ccaccgacgt gggcagaaag ctgatcatcg
     3961 accagaacgt gttcatcgag ggcaccctgc ctatgggagt cgtcagacca ctgaccgagg
     4021 tggaaatgga ccactacaga gagccctttc tgaaccccgt ggaccgggaa cctctttgga
     4081 gattccccaa cgagctgccc attgccggcg agcctgctaa tattgtggcc ctggtggaag
     4141 agtacatgga ctggctgcat cagagccccg tgcctaagct gctgttttgg ggaacacccg
     4201 gcgtgctgat tcctcctgct gaagctgcca gactggccaa gagcctgcct aattgcaagg
     4261 ccgtggatat cggccctggc ctgaatctgc tgcaagagga caaccccgat ctgatcggct
     4321 ccgagattgc cagatggctg agcaccctgg aaatcagcgg aggaggaggt tctggaggcg
     4381 gtggaagtgg tggcggaggt agcctgggct cgcgagccgc ggggttcgcg cggggcctgc
     4441 gggctgtggc act
//
