LOCUS       sequence-267765-        7323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human KIF3A 419-702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7323)
  AUTHORS   Yang R, Bostick Z, Garbouchian A, Luisi J, Banker G, Bentley M
  TITLE     A novel strategy to visualize vesicle-bound kinesins reveals the 
            diversity of kinesin-mediated transport.
  JOURNAL   Traffic. 2019 Aug 28. doi: 10.1111/tra.12692.
  PUBMED    31461551
REFERENCE   2  (bases 1 to 7323)
  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: "Traffic. 
            2019 Aug 28. doi: 10.1111/tra.12692."
COMMENT     Sequence Label: pBa.FRB-3myc-GS-KIF3A tail 419-702
FEATURES             Location/Qualifiers
     source          1..7323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..275
                     /label=chicken beta-actin promoter
     primer_bind     1218..1240
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1320..1598
                     /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"
     CDS             1605..1631
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2742..2876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2779..2798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2833..2852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2907..2923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2907..2923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2920..2942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2931..2947
                     /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(2955..2961,2962..2979,2980..2985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3000..3021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3138..3155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3309..3897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3389..3408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4068..4859,4860..4928))
                     /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     4691..4710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4929..5033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5101..5119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5157..5179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5279..5298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5492..5514
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5506..5523
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5507..5523
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggc
      121 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
      181 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      241 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      301 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      361 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      421 aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc cgggagggcc
      481 ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      541 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
      601 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
      661 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
      721 gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc cctccccgag ttgctgagca
      781 cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc gcggggctcg ccgtgccggg
      841 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg
      901 ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
      961 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1021 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     1081 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     1141 ccgtcccctt ctccatctcc agcctcgggg ctgtccgcag ggggacggct gccttcgggg
     1201 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggggttt atatcttccc
     1261 ttctctgttc ctccgcagcc cccaagcttg gcgcgcctag gcctaccatg gcttccagaa
     1321 tcctctggca tgagatgtgg catgaaggcc tggaagaggc atctcgtttg tactttgggg
     1381 aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt gcatgctatg atggaacggg
     1441 gcccccagac tctgaaggaa acatccttta atcaggccta tggtcgagat ttaatggagg
     1501 cccaagagtg gtgcaggaag tacatgaaat cagggaatgt caaggacctc ctccaagcct
     1561 gggacctcta ttatcatgtg ttccgacgaa tctcaaagac tagttatccg tacgacgtac
     1621 cagactacgc acagaagctg atctcagagg aggacctgca aaaacttatt tctgaagagg
     1681 atctgcaaaa acttatttct gaagaagatc tgggcggagg atccggaggc ggctctggcg
     1741 gagggctgta caagccagac aagatgattg aaatgcaagc aaaaattgat gaggagagaa
     1801 aagcacttga aacaaagctc gacatggaag aagaagaaag aaacaaggct agagctgaat
     1861 tagagaaacg ggaaaaagat cttcttaaag cccaacaaga gcatcagtct ttgctggaaa
     1921 aattatctgc cctggaaaag aaggtaattg ttggtggggt tgacttgttg gccaaagctg
     1981 aggaacaaga gaaacttctt gaagaatcta acatggaact ggaagaaagg aggaaaaggg
     2041 cagagcaact tcgcagagaa cttgaggaaa aagagcaaga acgcttggat attgaagaaa
     2101 aatataccag tttgcaagag gaagcacagg gaaagaccaa gaagttaaag aaagtttgga
     2161 ctatgctgat ggctgcaaag tcagagatgg ctgatctcca acaagaacat cagagggaaa
     2221 ttgaaggcct actggagaac attcggcaac ttagccggga gcttcgactt cagatgctta
     2281 ttattgataa ctttatacct cgggattatc aggaaatgat tgaaaactat gtccattgga
     2341 atgaagacat aggagaatgg cagctaaaat gtgttgctta tacaggaaat aacatgagga
     2401 agcaaacccc agtacctgat aaaaaggaga aagatccctt tgaggtggac ctttctcacg
     2461 tgtatcttgc ctatactgag gagagtctgc gtcagtcttt gatgaaacta gaaagaccac
     2521 gaacttcaaa ggggaaagca aggccaaaga cagggagaag aaagcgttct gcaaagcctg
     2581 aaactgtaat tgactctctg ctgcagtaac ggaccgtcta atctagagaa gtaatgatca
     2641 taatcagcca tatcacatct gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2701 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2761 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2821 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccccg
     2881 ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     2941 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3001 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3061 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3121 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3241 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5041 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5101 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     5161 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5221 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5281 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     5341 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     5401 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     5461 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     5521 agtgccaagt tgggatcttt gcattggccc acggctctca ggatggggat gctccccttc
     5581 agcacccggt tccccttgga aactgatggt cctggctctg tggcatggca gtggcactgt
     5641 gaggagcccc ctaccagcag cacacagtgg ggttggcact gccacgctcc ggatgccgcg
     5701 ctctgatcca accccataat caagggaacc cgaattgccc catcattgcc cccaccaccc
     5761 ccatcctgcc gggccctcac accccacgct gccttctggt gacattcccc agcccaaacc
     5821 cacggcttca tggctaccgc ggggcatttc ccattgccgc cccattatca gctctgcaca
     5881 cctcccgctg tacccatgcc tcgtggctgc ccttctttga cgtataatct tctaattaat
     5941 acccggcctt gtcaaagtgg agcacaaacg ttaattaatt ccccagcagg caggtaatta
     6001 acagtgtgac tccctttttg ctgcgagtgg ggctgataca gagagatgtg gcactatgga
     6061 gcccacgggg tcctggcact gggtgcccac ggaggtcccc atgtgctgca gtgtcaccgc
     6121 ctccgaggag acagtattgt ccctgcggtg tccctgcagc tcagctctgt ccacagggcc
     6181 acctccagtt tggaggggac acaatgcagc cccgatgcaa cccatcctcg cagcatccca
     6241 gggacaaaga ccccactgca agaccgcaca cagggctggg tcccgctccc ctaatatcta
     6301 cagtgcttct tgcatggccc cttaatcaat gcagttaatc agcatgcgct catgcaccgc
     6361 tctggagctg caaagcccct cgcagcgctg ctcaccaaca ccgcgcaccg ccccggccca
     6421 gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat tgcccaaatg taggcaaagg
     6481 cattcggctg ccttgacctc cgccgggccg ggccctgcct gactcagctc cttactcagc
     6541 gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca ccctgacaaa gagtggccct
     6601 taacgggctc tgaggtgcac ccagcagtgc actcagcagt ccaagggccg gcctggaggt
     6661 ttgcaccgct acgtgctgac attagcattg aacttggccc tgggtagtgc tgcaggccgg
     6721 gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg tgccccttcc cctcccttgc
     6781 atcccagcag gctgcacccc agcaccaggc ccgtgcatgc atgctcctgg tgttattgca
     6841 gcctggtgca tgcatgcgtc ttgatggtgc agcgctgtgc atgcatcctc cttggtgtgt
     6901 agcagtctag tgcatgcata cccctcggtg ttattgctgc tctgtgcacg cacgctcatt
     6961 gtatcacttc atcccagtgc atgcactcac actggagcga ttgctgctcg gtgcacgcac
     7021 actcattgta tcactgcagc tcagtggctg cacgcacacc ggtgttattg ctgctcggtg
     7081 cgtgcatgca catcagtgtc gctgcagctc agtgcatgca cgctcattgc ccatcgctat
     7141 ccctgcctct cctgctggcg ctccccggga ggtgacttca aggggaccgc aggaccacct
     7201 cgggggtggg gggagggctg cacacgcgga ccccgctccc cctccccaac aaagcactgt
     7261 ggaatcaaaa aggggggagg ggggatggag gggcgcgtca cacccccgcc ccacaccctc
     7321 acc
//
