LOCUS       sequence-349333-        4453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human KCNC3 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: KCNC3_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..897
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(1381..1398)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1381..1397)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1390..1412)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1606..1625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1725..1747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1785..1803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1871..1975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1976..2044,2045..2836)
                     /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     complement(2194..2213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3007..3595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3496..3515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3749..3766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3883..3904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3919..3924,3925..3942,3943..3949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3957..3973
                     /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)."
     primer_bind     3962..3984
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3981..3997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3981..3997
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgtccaatgg ccgagatcgg caccggcttt cccttcgatc ctcactacgt ggaagtgctg
       61 ggcgagagaa tgcactatgt ggacgtgggc cccagagatg gaacccctgt gctgtttctg
      121 cacggcaacc ctaccagcag ctacgtgtgg cggaacatca tccctcacgt ggcccctaca
      181 cacagatgta tcgcccctga cctgatcggc atgggcaaga gcgataagcc cgacctgggc
      241 tacttcttcg acgaccacgt gcggttcatg gacgccttta ttgaggccct cggcctggaa
      301 gaggtggtgc tggttattca cgattggggc agcgccctgg gattccactg ggccaagaga
      361 aaccccgaga gagtgaaggg aatcgccttc atggaattca tcaggcccat tcctacctgg
      421 gacgagtggc ctgagttcgc cagagagaca ttccaggcct tcagaaccac cgacgtgggc
      481 agaaagctga tcatcgacca gaacgtgttc atcgagggca ccctgcctat gggagtcgtc
      541 agaccactga ccgaggtgga aatggaccac tacagagagc cctttctgaa ccccgtggac
      601 cgggaacctc tttggagatt ccccaacgag ctgcccattg ccggcgagcc tgctaatatt
      661 gtggccctgg tggaagagta catggactgg ctgcatcaga gccccgtgcc taagctgctg
      721 ttttggggaa cacccggcgt gctgattcct cctgctgaag ctgccagact ggccaagagc
      781 ctgcctaatt gcaaggccgt ggatatcggc cctggcctga atctgctgca agaggacaac
      841 cccgatctga tcggctccga gattgccaga tggctgagca ccctggaaat cagcggagga
      901 ggaggttctg gaggcggtgg aagtggtggc ggaggtagcc tgagctcagt ctgcgtctcg
      961 tccttccgcg ggcgccaggg ggccagcaag cagcagccgg cgccaccgcc gcagccgccc
     1021 gagtccccgc cgccgccacc gctgccgccg cagcagcagc agcctgcgca gcccggcccc
     1081 gccgcgtccc cggcgggccc cccggcaccc cgcgggcccg gggaccggcg cgccgagcca
     1141 tgccccgggc tgccggcggc ggccatgggg cggcacggcg gcggcggtgg cgacagcggc
     1201 aagatcgtga tcaacgtggg cggcgtgcgc catgagacgt accgctcgac gctgcgcacc
     1261 ctgccgggga cgcggctggc cggcctgacg gagcccgagg cggcggcacg cttcgactac
     1321 gacccgggcg ccgacgagtc caatatctag atgcattcgc gaggtaccga gctcgaattc
     1381 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1441 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     1501 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     1561 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     1621 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1741 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg
     2701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3781 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3841 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     3901 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     3961 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg
     4021 caggcctctg cagtcgacgg gcccgggatc cgatccaatc cctctacccc gcccgcactt
     4081 ttttcagttc agtaaggtct ctctctatcg tatctagccc cgccttcccc tggctaccag
     4141 tgctgccgaa cacaattagt agtaaccggt cccgcctccc ttgacgcccc gccccgcccc
     4201 tcctccctgc tctcttagaa agagccaatc gctccgtctc attggcggcc ccgactcgtc
     4261 cagctgggtc gcgttagagt gcccgcccca ctcctcccag ccacagcccc gccccacctc
     4321 ccccaggcca atcagtttgg ttcctctccc ctaagccacg cccccgctac ctcgctcctc
     4381 ctcccaccca atcccgtcgg tctctccccc cgtgtccccg cccctgcgtc cccgcccctc
     4441 ccgccccgcc ccc
//
