LOCUS       sequence-426209-        8941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Imaging of KIF5B fused to HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8941)
  TITLE     Imaging of KIF5B and Tubulin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8941)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: hsKIF5B(1-560)-HaloTag-FRB
FEATURES             Location/Qualifiers
     source          1..8941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1702..2586
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             2593..2871
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     polyA_signal    3070..3204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3107..3126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3161..3180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3218..3234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3218..3234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3231..3253)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3242..3258
                     /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(3266..3272,3273..3290,3291..3296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3311..3332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3449..3466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3620..4208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3700..3719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4379..5170,5171..5239))
                     /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     5002..5021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5240..5344)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5412..5430
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5468..5490)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5590..5609
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5803..5825
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5817..5834
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5818..5834
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7635..7910
                     /label=chicken beta-actin promoter
     primer_bind     8853..8875
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      join(8936..8941,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggcgcgc ctatggcgga cctggccgag tgcaacatca aagtgatgtg tcgcttcaga
       61 cctctcaacg agtctgaagt gaaccgcggc gacaagtaca tcgccaagtt tcagggagaa
      121 gacacggtcg tgatcgcgtc caagccttat gcatttgatc gggtgttcca gtcaagcaca
      181 tctcaagagc aagtgtataa tgactgtgca aagaagattg ttaaagatgt acttgaagga
      241 tataatggaa caatatttgc atatggacaa acatcctctg ggaagacaca cacaatggag
      301 ggtaaacttc atgatccaga aggcatggga attattccaa gaatagtgca agatattttt
      361 aattatattt actccatgga tgaaaatttg gaatttcata ttaaggtttc atattttgaa
      421 atatatttgg ataagataag ggacctgtta gatgtttcaa agaccaacct ttcagttcat
      481 gaagacaaaa accgagttcc ctatgtaaag gggtgcacag agcgttttgt atgtagtcca
      541 gatgaagtta tggataccat agatgaagga aaatccaaca gacatgtagc agttacaaat
      601 atgaatgaac atagctctag gagtcacagt atatttctta ttaatgtcaa acaagagaac
      661 acacaaacgg aacaaaagct gagtggaaaa ctttatctgg ttgatttagc tggtagtgaa
      721 aaggttagta aaactggagc tgaaggtgct gtgctggatg aagctaaaaa catcaacaag
      781 tcactttctg ctcttggaaa tgttatttct gctttggctg agggtagtac atatgttcca
      841 tatcgagata gtaaaatgac aagaatcctt caagattcat taggtggcaa ctgtagaacc
      901 actattgtaa tttgctgctc tccatcatca tacaatgagt ctgaaacaaa atctacactc
      961 ttatttggcc aaagggccaa aacaattaag aacacagttt gtgtcaatgt ggagttaact
     1021 gcagaacagt ggaaaaagaa gtatgaaaaa gaaaaagaaa aaaataagat cctgcggaac
     1081 actattcagt ggcttgaaaa tgagctcaac agatggcgta atggggagac ggtgcctatt
     1141 gatgaacagt ttgacaaaga gaaagccaac ttggaagctt tcacagtgga taaagatatt
     1201 actcttacca atgataaacc agcaaccgca attggagtta taggaaattt tactgatgct
     1261 gaaagaagaa agtgtgaaga agaaattgct aaattataca aacagcttga tgacaaggat
     1321 gaagaaatta accagcaaag tcaactggta gagaaactga agacgcaaat gttggatcag
     1381 gaggagcttt tggcatctac cagaagggat caagacaata tgcaagctga gctgaatcgc
     1441 cttcaagcag aaaatgatgc ctctaaagaa gaagtgaaag aagttttaca ggccctagaa
     1501 gaacttgctg tcaattatga tcagaagtct caggaagttg aagacaaaac taaggaatat
     1561 gaattgctta gtgatgaatt gaatcagaaa tcggcaactt tagcgagtat agatgctgag
     1621 cttcagaaac ttaaggaaat gaccaaccac cagaaaaaac gagcagctga gatgatggca
     1681 tctttactaa aaaagtcgac tgaaatcggt actggctttc cattcgaccc ccattatgtg
     1741 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
     1801 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
     1861 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
     1921 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
     1981 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
     2041 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
     2101 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
     2161 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
     2221 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
     2281 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
     2341 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
     2401 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
     2461 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
     2521 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtccac gctcgagatt
     2581 tccggcacta gtatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt
     2641 ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct
     2701 atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga
     2761 gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac
     2821 ctcctccaag cctgggacct ctattatcat gtgttccgac gaatctcaaa gtaagcggcc
     2881 gccatatgca tcctaggcct attaatattc cggagtatac gtagccggct aacgttaaca
     2941 accggtacct ctagataagt aatgatcata atcagccata tcacatctgt agaggtttta
     3001 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3061 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3181 aatgtatctt atcatgtctg gatcaattcg taatcatgtc atagctgttt cctgtgtgaa
     3241 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3301 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3361 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4141 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4201 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4261 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4321 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4381 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4441 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4501 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4561 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4621 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4681 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4741 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4801 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4861 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4921 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4981 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5041 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5101 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5161 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5221 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5281 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5341 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5401 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     5461 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5521 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     5581 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     5641 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     5701 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5761 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     5821 aaacgacggc cagtgccaag ttgggatctt tgcattggcc cacggctctc aggatgggga
     5881 tgctcccctt cagcacccgg ttccccttgg aaactgatgg tcctggctct gtggcatggc
     5941 agtggcactg tgaggagccc cctaccagca gcacacagtg gggttggcac tgccacgctc
     6001 cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc ccatcattgc
     6061 ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttctgg tgacattccc
     6121 cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg ccccattatc
     6181 agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg acgtataatc
     6241 ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat tccccagcag
     6301 gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac agagagatgt
     6361 ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc catgtgctgc
     6421 agtgtcaccg cctccgagga gacagtattg tccctgcggt gtccctgcag ctcagctctg
     6481 tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca acccatcctc
     6541 gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg gtcccgctcc
     6601 cctaatatct acagtgcttc ttgcatggcc ccttaatcaa tgcagttaat cagcatgcgc
     6661 tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct gctcaccaac accgcgcacc
     6721 gccccggccc agcctgcagc acgcgctgca aacaggaaag aaacaaaata ttgcccaaat
     6781 gtaggcaaag gcattcggct gccttgacct ccgccgggcc gggccctgcc tgactcagct
     6841 ccttactcag cgctcgcttc ctccctccgg ctgccaccgc cgcagcgcac accctgacaa
     6901 agagtggccc ttaacgggct ctgaggtgca cccagcagtg cactcagcag tccaagggcc
     6961 ggcctggagg tttgcaccgc tacgtgctga cattagcatt gaacttggcc ctgggtagtg
     7021 ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc gttgcacccg gtgccccttc
     7081 ccctcccttg catcccagca ggctgcaccc cagcaccagg cccgtgcatg catgctcctg
     7141 gtgttattgc agcctggtgc atgcatgcgt cttgatggtg cagcgctgtg catgcatcct
     7201 ccttggtgtg tagcagtcta gtgcatgcat acccctcggt gttattgctg ctctgtgcac
     7261 gcacgctcat tgtatcactt catcccagtg catgcactca cactggagcg attgctgctc
     7321 ggtgcacgca cactcattgt atcactgcag ctcagtggct gcacgcacac cggtgttatt
     7381 gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct cagtgcatgc acgctcattg
     7441 cccatcgcta tccctgcctc tcctgctggc gctccccggg aggtgacttc aaggggaccg
     7501 caggaccacc tcgggggtgg ggggagggct gcacacgcgg accccgctcc ccctccccaa
     7561 caaagcactg tggaatcaaa aaggggggag gggggatgga ggggcgcgtc acacccccgc
     7621 cccacaccct cacctcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     7681 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     7741 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
     7801 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
     7861 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
     7921 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
     7981 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
     8041 attagcgctt ggtttaatga cggctcgttt cttttctgtg gctgcgtgaa agccttaaag
     8101 ggctccggga gggccctttg tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     8161 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     8221 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     8281 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     8341 gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc acccccctcc
     8401 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg
     8461 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
     8521 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
     8581 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     8641 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     8701 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     8761 tgcgtcgccg cgccgccgtc cccttctcca tctccagcct cggggctgtc cgcaggggga
     8821 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     8881 ggtttatatc ttcccttctc tgttcctccg cagcccccaa gctagcgcta agcttgccac
     8941 c
//
