LOCUS       sequence-349353-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human VCP at the C-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4442)
  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: VCP_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..44)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(198..786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(278..297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(957..1748,1749..1817))
                     /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     1580..1599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1818..1922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1990..2008
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2046..2068)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2168..2187
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2381..2403
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2395..2412
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2396..2412
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2893..3780
                     /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(4238..4254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4238..4254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4251..4273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4262..4278
                     /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(4286..4292,4293..4310,4311..4316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4331..4352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
       61 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1081 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag
     1141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1921 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     1981 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2041 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2101 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2161 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2221 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     2281 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2341 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2401 acgacggcca gtgaattcga gctcggtacc tcgcgaatgc atctagagga tgatccagtg
     2461 cctgagatcc gtcgagatca ctttgaagaa gccatgcgct ttgcgcgccg ttctgtcagt
     2521 gacaatgaca ttcggaagta tgagatgttt gcccagaccc ttcagcagag tcggggcttt
     2581 ggcagcttca ggtaagttgg ttgggagcat tagacagtgc ttaagttact ttggggacct
     2641 acaccaaaag ggatgggagt cctaaggaag ctagaggggt agttgtggaa atcttacaca
     2701 ggccctgtcc taaccctctt ttttggcttt gctcttgtac acagattccc ttcagggaac
     2761 cagggtggag ctggccccag tcagggcagt ggaggcggca caggtggcag tgtatacaca
     2821 gaagacaatg atgatgacct gtatggcggt ggaggaggtt ctggaggcgg tggaagtggt
     2881 ggcggaggta gcgccgagat cggcaccggc tttcccttcg atcctcacta cgtggaagtg
     2941 ctgggcgaga gaatgcacta tgtggacgtg ggccccagag atggaacccc tgtgctgttt
     3001 ctgcacggca accctaccag cagctacgtg tggcggaaca tcatccctca cgtggcccct
     3061 acacacagat gtatcgcccc tgacctgatc ggcatgggca agagcgataa gcccgacctg
     3121 ggctacttct tcgacgacca cgtgcggttc atggacgcct ttattgaggc cctcggcctg
     3181 gaagaggtgg tgctggttat tcacgattgg ggcagcgccc tgggattcca ctgggccaag
     3241 agaaaccccg agagagtgaa gggaatcgcc ttcatggaat tcatcaggcc cattcctacc
     3301 tgggacgagt ggcctgagtt cgccagagag acattccagg ccttcagaac caccgacgtg
     3361 ggcagaaagc tgatcatcga ccagaacgtg ttcatcgagg gcaccctgcc tatgggagtc
     3421 gtcagaccac tgaccgaggt ggaaatggac cactacagag agccctttct gaaccccgtg
     3481 gaccgggaac ctctttggag attccccaac gagctgccca ttgccggcga gcctgctaat
     3541 attgtggccc tggtggaaga gtacatggac tggctgcatc agagccccgt gcctaagctg
     3601 ctgttttggg gaacacccgg cgtgctgatt cctcctgctg aagctgccag actggccaag
     3661 agcctgccta attgcaaggc cgtggatatc ggccctggcc tgaatctgct gcaagaggac
     3721 aaccccgatc tgatcggctc cgagattgcc agatggctga gcaccctgga aatcagcgga
     3781 taagtggtgg tggccagcgt gcagtgagct ggcctgcctg gaccttgttc cctgggggtg
     3841 ggggcgcttg cccaggagag ggaccagggg tgcgcccaca gcctgctcca ttctccagtc
     3901 tgaacagttc agctacagtc tgactctgga cagggggttt ctgttgcaaa aatacaaaac
     3961 aaaagcgata aaataaaagc gattttcatt tggtaggcgg agagtgaatt accaacaggg
     4021 aattgggcct tgggcctatg ccatttctgt tgtagtttgg ggcagtgcag gggacctgtg
     4081 tggggtgtga accaaggcac tactgccacc tgccacagta aagcatctgc acttgactca
     4141 atgctgcccg agccctccct tccccctatc caacctgggt aggggatccc gggcccgtcg
     4201 actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4261 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4321 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4381 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4441 gt
//
