LOCUS       sequence-349373-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PRKCG 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: PRKCG_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     331..353
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     345..362
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     346..362
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             843..1730
                     /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(2188..2204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2188..2204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2201..2223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2212..2228
                     /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(2236..2242,2243..2260,2261..2266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2281..2302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2419..2436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2590..3178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2670..2689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3349..4140,4141..4209))
                     /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     3972..3991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4210..4314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4382..4400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4438..4442,1..18))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
       61 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
      121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
      181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
      241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      361 gtgaattcga gctcggtacc tcgcgaatgc atctagatga aggggtgggg ttcccctggg
      421 cctcaatata cctgtatgtg ggggtggggt tccctctgca gagccccccg cccccaacaa
      481 aaggaggtgc agacaccatg aagcatgaat agagattctg caggagacag gagatgagac
      541 tggggtacac agagggacac ccgaggagcc ctcggagctg cttaactttc cctcccccac
      601 gtctcccaca gtgtggccgc agcggcgaga actttgacaa gttcttcacg cgggcggcgc
      661 cagcgctgac ccctccagac cgcctagtcc tggccagcat cgaccaggcc gatttccagg
      721 gcttcaccta cgtgaacccc gacttcgtgc acccggatgc ccgcagcccc accagcccag
      781 tgcctgtgcc cgtcatgggt ggaggaggtt ctggaggcgg tggaagtggt ggcggaggta
      841 gcgccgagat cggcaccggc tttcccttcg atcctcacta cgtggaagtg ctgggcgaga
      901 gaatgcacta tgtggacgtg ggccccagag atggaacccc tgtgctgttt ctgcacggca
      961 accctaccag cagctacgtg tggcggaaca tcatccctca cgtggcccct acacacagat
     1021 gtatcgcccc tgacctgatc ggcatgggca agagcgataa gcccgacctg ggctacttct
     1081 tcgacgacca cgtgcggttc atggacgcct ttattgaggc cctcggcctg gaagaggtgg
     1141 tgctggttat tcacgattgg ggcagcgccc tgggattcca ctgggccaag agaaaccccg
     1201 agagagtgaa gggaatcgcc ttcatggaat tcatcaggcc cattcctacc tgggacgagt
     1261 ggcctgagtt cgccagagag acattccagg ccttcagaac caccgacgtg ggcagaaagc
     1321 tgatcatcga ccagaacgtg ttcatcgagg gcaccctgcc tatgggagtc gtcagaccac
     1381 tgaccgaggt ggaaatggac cactacagag agccctttct gaaccccgtg gaccgggaac
     1441 ctctttggag attccccaac gagctgccca ttgccggcga gcctgctaat attgtggccc
     1501 tggtggaaga gtacatggac tggctgcatc agagccccgt gcctaagctg ctgttttggg
     1561 gaacacccgg cgtgctgatt cctcctgctg aagctgccag actggccaag agcctgccta
     1621 attgcaaggc cgtggatatc ggccctggcc tgaatctgct gcaagaggac aaccccgatc
     1681 tgatcggctc cgagattgcc agatggctga gcaccctgga aatcagcgga taatctcacc
     1741 cgccgccact aggtgtcccc aacgtcccct ccgccgtgcc ggcggcagcc ccacttcacc
     1801 cccaacttca ccaccccctg tcccattcta gatcctgcac cccagcattc cagctctgcc
     1861 cccgcgggtt ctagacgccc ctcccaagcg ttcctggcct tctgaactcc atacagcctc
     1921 tacagccgtc ccgcgttcaa gacttgagcg gagcccgata ttctccctga ccttagcgtt
     1981 ctggactctg ccccaatcgg gtccagagac cacaccacta accatcccca actccatggg
     2041 gttcgagact ccatcttggt agttctgtgc ctccccccag accccgcccc tggggaaata
     2101 gcctcacggg gttggctgtt ccagactcag gttggatccc gggcccgtcg actgcagagg
     2161 cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2221 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2281 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2341 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2401 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2461 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2521 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2581 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2641 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2701 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2761 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2821 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2881 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2941 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3001 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3061 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3121 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3181 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3241 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3301 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3421 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3481 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3541 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3781 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3841 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4021 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4201 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4321 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4381 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4441 ct
//
