LOCUS       sequence-349370-        4380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human CACNA1A at the 
            N-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4380)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4380)
  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: CACNA1A_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             289..1179
                     /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(1646..1662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1646..1662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1659..1681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1670..1686
                     /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(1694..1700,1701..1718,1719..1724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1739..1760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1877..1894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2048..2636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2128..2147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2807..3598,3599..3667))
                     /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     3430..3449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3668..3772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3840..3858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3896..3918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4018..4037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4231..4253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4245..4262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4246..4262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccccgccccc gcaccctcct cccgcccctc cttctccggg gtcagccagg aagatgtccc
       61 gagctgctat ccccggctcg gcccgggcag ccgccttctg agcccccgac ccgaggcgcc
      121 gagccgccgc cgcccgatgg gctgggccgt ggagcgtctc cgcagtcgta gctccagccg
      181 ccgcgctccc agccccggca gcctcagcat cagcggcggc ggcggcggcg gcggcgtctt
      241 ccgcatcgtt cgccgcagcg taacccggag ccctttgctc tttgcagaat ggccgagatc
      301 ggcaccggct ttcccttcga tcctcactac gtggaagtgc tgggcgagag aatgcactat
      361 gtggacgtgg gccccagaga tggaacccct gtgctgtttc tgcacggcaa ccctaccagc
      421 agctacgtgt ggcggaacat catccctcac gtggccccta cacacagatg tatcgcccct
      481 gacctgatcg gcatgggcaa gagcgataag cccgacctgg gctacttctt cgacgaccac
      541 gtgcggttca tggacgcctt tattgaggcc ctcggcctgg aagaggtggt gctggttatt
      601 cacgattggg gcagcgccct gggattccac tgggccaaga gaaaccccga gagagtgaag
      661 ggaatcgcct tcatggaatt catcaggccc attcctacct gggacgagtg gcctgagttc
      721 gccagagaga cattccaggc cttcagaacc accgacgtgg gcagaaagct gatcatcgac
      781 cagaacgtgt tcatcgaggg caccctgcct atgggagtcg tcagaccact gaccgaggtg
      841 gaaatggacc actacagaga gccctttctg aaccccgtgg accgggaacc tctttggaga
      901 ttccccaacg agctgcccat tgccggcgag cctgctaata ttgtggccct ggtggaagag
      961 tacatggact ggctgcatca gagccccgtg cctaagctgc tgttttgggg aacacccggc
     1021 gtgctgattc ctcctgctga agctgccaga ctggccaaga gcctgcctaa ttgcaaggcc
     1081 gtggatatcg gccctggcct gaatctgctg caagaggaca accccgatct gatcggctcc
     1141 gagattgcca gatggctgag caccctggaa atcagcggag gaggaggttc tggaggcggt
     1201 ggaagtggtg gcggaggtag cgcccgcttc ggagacgaga tgccggcccg ctacggggga
     1261 ggaggctccg gggcagccgc cggggtggtc gtgggcagcg gaggcgggcg aggagccggg
     1321 ggcagccggc agggcgggca gcccggggcg caaaggatgt acaagcagtc aatggcgcag
     1381 agagcgcgga ccatggcact ctacaacccc atccccgtcc gacagaactg cctcacggtt
     1441 aaccggtctc tcttcctctt cagcgaagac aacgtggtga gaaaatacgc caaaaagatc
     1501 accgaatggc catatccttt tgcccgaacc ccagcagcag ctgcgcctcc ccctcctccc
     1561 tccgcctccc ctcttccagg ctgggagaga gacccggggg ttgatgggag gtggggagga
     1621 gctcgagaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1681 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1741 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2221 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2281 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2341 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2401 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2461 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2521 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2581 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2641 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2701 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2761 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2941 taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3661 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3781 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     3841 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     3901 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3961 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4021 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4081 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     4141 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4201 ggcgattaag ttgggtaacg ccagggttta cccagtcacg acgttgtaaa acgacggcca
     4261 gtgaattcct cttttttttt ttttccggag tgggtggact ggggattgcc tggattcctt
     4321 cctcggttat tttttgcctc gcttctctct cccctccccc ctcctccccg ggcccccgcg
//
