LOCUS       sequence-349380-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human KIF5A 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: KIF5A_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2048..2070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2170..2189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2383..2405
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2397..2414
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2398..2414
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2866..3753
                     /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(4175..4191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4175..4191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4188..4210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4199..4215
                     /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(4223..4229,4230..4247,4248..4253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4268..4289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     2221 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     2281 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     2341 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     2401 aaacgacggc cagtgaattc gagatccagt cattcccctg cctcagtgca gaatatctcc
     2461 cctcctccaa tcccatctgt ggcttctccc tattccagtg ataaggtctg gacgaaaaca
     2521 acgtgggcat ctgacgactg ggtttctctc ctacctttgg aagccttgtg acattggaca
     2581 agtcatttac atccctctgg ccttgagttt ccctgtattc ctgtaaaaca caaatagtcc
     2641 ctgccgtttg tgatctcaca gggacactct cagagcagag taaccatgat gacaggaata
     2701 ctttaatttc tatggagctg atcatggtgg gtctcttcct ccaggagtga cctgccgtgt
     2761 ggctatgagg ctgaggacca ggccaagctt ttccctctcc accaagagac agcagccagc
     2821 ggtggaggag gttctggagg cggtggaagt ggtggcggag gtagcgccga gatcggcacc
     2881 ggctttccct tcgatcctca ctacgtggaa gtgctgggcg agagaatgca ctatgtggac
     2941 gtgggcccca gagatggaac ccctgtgctg tttctgcacg gcaaccctac cagcagctac
     3001 gtgtggcgga acatcatccc tcacgtggcc cctacacaca gatgtatcgc ccctgacctg
     3061 atcggcatgg gcaagagcga taagcccgac ctgggctact tcttcgacga ccacgtgcgg
     3121 ttcatggacg cctttattga ggccctcggc ctggaagagg tggtgctggt tattcacgat
     3181 tggggcagcg ccctgggatt ccactgggcc aagagaaacc ccgagagagt gaagggaatc
     3241 gccttcatgg aattcatcag gcccattcct acctgggacg agtggcctga gttcgccaga
     3301 gagacattcc aggccttcag aaccaccgac gtgggcagaa agctgatcat cgaccagaac
     3361 gtgttcatcg agggcaccct gcctatggga gtcgtcagac cactgaccga ggtggaaatg
     3421 gaccactaca gagagccctt tctgaacccc gtggaccggg aacctctttg gagattcccc
     3481 aacgagctgc ccattgccgg cgagcctgct aatattgtgg ccctggtgga agagtacatg
     3541 gactggctgc atcagagccc cgtgcctaag ctgctgtttt ggggaacacc cggcgtgctg
     3601 attcctcctg ctgaagctgc cagactggcc aagagcctgc ctaattgcaa ggccgtggat
     3661 atcggccctg gcctgaatct gctgcaagag gacaaccccg atctgatcgg ctccgagatt
     3721 gccagatggc tgagcaccct ggaaatcagc ggataatctc ccacacccac ggctgcatac
     3781 ctgcactttc aggtagcgtc aggctgcttc ctcggaccag cctcaggttg cttcccttct
     3841 tgctgacagc ctctttggtt ttcctgctgc tgcttctttt tttttttatc acctcaaaac
     3901 tttaattatg tagttacccc tcaactcctg ctttcccctg ttgggggcta cacctgtgct
     3961 gccccatgta atctggacgt tgacctcccg tgcccccttt tgctttccgt taggctacag
     4021 tggtaaccac agtactgtct ttttgccact aggggtcact gtcacatgtc caaagaagtg
     4081 agcttggccg ctgtagatta gaaccacagg ggtggtagac tgcagatcta tcaggagacc
     4141 ctagggggcc atctcaaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4201 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4261 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4321 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcg
//
