LOCUS       sequence-349081-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human KCND3 at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4377)
  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: KCND3_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(262..850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(342..361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1021..1812,1813..1881))
                     /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     1644..1663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1882..1986)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2054..2072
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2110..2132)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2232..2251
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2445..2467
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2459..2476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2460..2476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2928..3815
                     /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(4237..4253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4237..4253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4250..4272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4261..4277
                     /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(4285..4291,4292..4309,4310..4315))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4330..4351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
       61 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      121 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1141 cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa
     1201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1981 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     2041 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     2101 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2161 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     2221 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     2281 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     2341 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2401 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2461 taaaacgacg gccagtgaat tcctactata tacaacttgg taatttctgg tagtttggga
     2521 ataataatta gaatggataa ttacaagttg attacaaaag gcagagatag aggcacaaca
     2581 aaatatcctc ctagttacca cgagcaagaa atgtaggggg tcaaggaagc tttgggacgc
     2641 tggccagcag gaaccatcat caactcactc ttttctatcc cagtcgctcc agccttaatt
     2701 tgaaagcaga cgacggactg agaccaaact gcaaaacatc ccagatcacc acagccatca
     2761 tcagcatccc cactccccca gcgctaaccc cagaggggga aagtcggcca ccccctgcca
     2821 gcccaggccc caacacgaac attccttcca tagccagcaa tgttgtcaag gtctccgcct
     2881 tgggtggagg aggttctgga ggcggtggaa gtggtggcgg aggtagcgcc gagatcggca
     2941 ccggctttcc cttcgatcct cactacgtgg aagtgctggg cgagagaatg cactatgtgg
     3001 acgtgggccc cagagatgga acccctgtgc tgtttctgca cggcaaccct accagcagct
     3061 acgtgtggcg gaacatcatc cctcacgtgg cccctacaca cagatgtatc gcccctgacc
     3121 tgatcggcat gggcaagagc gataagcccg acctgggcta cttcttcgac gaccacgtgc
     3181 ggttcatgga cgcctttatt gaggccctcg gcctggaaga ggtggtgctg gttattcacg
     3241 attggggcag cgccctggga ttccactggg ccaagagaaa ccccgagaga gtgaagggaa
     3301 tcgccttcat ggaattcatc aggcccattc ctacctggga cgagtggcct gagttcgcca
     3361 gagagacatt ccaggccttc agaaccaccg acgtgggcag aaagctgatc atcgaccaga
     3421 acgtgttcat cgagggcacc ctgcctatgg gagtcgtcag accactgacc gaggtggaaa
     3481 tggaccacta cagagagccc tttctgaacc ccgtggaccg ggaacctctt tggagattcc
     3541 ccaacgagct gcccattgcc ggcgagcctg ctaatattgt ggccctggtg gaagagtaca
     3601 tggactggct gcatcagagc cccgtgccta agctgctgtt ttggggaaca cccggcgtgc
     3661 tgattcctcc tgctgaagct gccagactgg ccaagagcct gcctaattgc aaggccgtgg
     3721 atatcggccc tggcctgaat ctgctgcaag aggacaaccc cgatctgatc ggctccgaga
     3781 ttgccagatg gctgagcacc ctggaaatca gcggataaaa ccactggaca gagggccaga
     3841 gtgggtagtg gggaatgaag gggactggca tgttggtggg tggtcactga gaccactccc
     3901 tccccccttt ccccactatt tctgcctgcc ccattgtacc cctagcactg agacttgtgc
     3961 ctggaaggaa aaagaggtag caaaggggca cctgaggttc acgctgctag cgtggacata
     4021 gccctgtgat actgcatctc actggggcca gaggaaggga gggagggtgg gggggtcggg
     4081 gataggccgg gggatatagg ctatgtgcca ggacagggcc tggatggagg agcctcagac
     4141 tcagaccaaa taacagtcgt ttctggagac cccagtgagg aaaatacaag accaagagca
     4201 gcaaagaggc caagttgtaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4261 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4321 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgcttt
//
