LOCUS       sequence-232540-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active Kinesin-3 motor tagged with DHFR and myc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Gutnick A, Banghart MR, West ER, Schwarz TL
  TITLE     The light-sensitive dimerizer zapalog reveals distinct modes of 
            immobilization for axonal mitochondria.
  JOURNAL   Nat Cell Biol. 2019 Jun;21(6):768-777. doi: 
            10.1038/s41556-019-0317-2. Epub 2019 May 6.
  PUBMED    31061466
REFERENCE   2  (bases 1 to 6944)
  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"; doi: 
            "10.1038/s41556-019-0317-2"; journalName: "Nat Cell Biol"; date: 
            "2019-06"; volume: "21"; issue: "6"; pages: "768-777"
COMMENT     Sequence Label: Kif1A(1-489)-DHFR-myc
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1911..1920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2414..2887
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2888..2917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2952..2969)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2958..3182
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3228..3656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3315..3334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3525..3546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3665..3685)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3670..3999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3850..3985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3912..3931
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4047..4094
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4113..4487
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4617..4738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4654..4673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4708..4727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4787..4803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4787..4803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4800..4822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4811..4827
                     /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(4835..4841,4842..4859,4860..4865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4880..4901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5018..5035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5269..5288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5948..6739,6740..6808))
                     /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     6571..6590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6809..6913)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatggct ggggcctctg tgaaggtggc
      961 ggtgcgcgtc cgccccttca actcccggga aatgagccgc gactccaagt gcatcattca
     1021 gatgtctgga aacaccacca ccattgtcaa ccccaaacag cccaaggaga cacccaagag
     1081 cttcagcttc gattactcct actggtcaca tacctcgccc gaggacatca actatgcctc
     1141 tcagaagcag gtctacaggg acattgggga ggagatgcta cagcatgcct ttgagggcta
     1201 caacgtgtgc atctttgcct atggacagac aggggctggc aagtcctaca ccatgatggg
     1261 caaacaagag aaggaccagc aaggcatcat cccacagctc tgtgaagacc tcttctctcg
     1321 aatcaatgac acgaccaacg acaacatgtc ctattctgtg gaggtcagct acatggagat
     1381 ttactgtgag cgagtccgcg acctcctgaa ccccaagaac aaaggcaacc tgcgtgtgag
     1441 ggggcaccct ctgctggggc cttatgtgga ggacctctcc aagctggctg tgacctccta
     1501 caatgacatc caggacctca tggactcagg gaacaaggcc aggactgtgg cggccacaaa
     1561 catgaatgag accagcagcc gttctcacgc cgtcttcaac atcatcttca cccagaagcg
     1621 ccatgatgcg gagaccaaca tcaccacaga gaaagtgagc aaaatcagcc tggtggacct
     1681 ggcgggcagt gaacgagccg actccacggg agccaagggc acacgactca aggagggagc
     1741 caacatcaac aagtctttga ccaccctggg aaaggtcatc tctgctctcg ctgaaatgga
     1801 ctctggaccc aataagaaca agaaaaagaa gaagacagac ttcatccctt accgagactc
     1861 ggtattgaca tggcttctcc gggaaaacct gggcggcaac tcaaggacag ccatggtggc
     1921 agccctgagc cctgcggaca tcaactatga tgagaccctc agcacactga gatatgcaga
     1981 ccgggccaaa cagatccgct gcaacgccat tatcaatgag gaccctaaca acaagctgat
     2041 tcgagaactg aaggatgagg tgaccaggct gcgagatctg ctgtacgccc aaggtcttgg
     2101 cgatatcact gacaccaaca ctgtgcctgg aggacccaaa ttgaccaatg ccctggtggg
     2161 catgagcccc tcgtcttcgc tctcagccct gtccagccgt gcggcctctg tgtccagtct
     2221 ccacgagcgc atcttgtttg ctccaggcag cgaggaggct attgaaaggc tgaaggaaac
     2281 agagaagatc attgctgagc tcaatgagac gtgggaggag aagctacggc ggacagaagc
     2341 catccggatg gagagggaag ccctgctggc cgagatgggt gtggccatga gggaagatgg
     2401 tctcgagtct agaatcagtc tgattgcggc gttagcggta gatcgcgtta tcggcatgga
     2461 aaacgccatg ccgtggaacc tgcctgccga tctcgcctgg tttaaacgca acaccttaaa
     2521 taaacccgtg attatgggcc gccatacctg ggaatcaatc ggtcgtccgt tgccaggacg
     2581 caaaaatatt atcctcagca gtcaaccggg tacggacgat cgcgtaacgt gggtgaagtc
     2641 ggtggatgaa gccatcgcgg cgtgtggtga cgtaccagaa atcatggtga ttggcggcgg
     2701 tcgcgtttat gaacagttct tgccaaaagc gcaaaaactg tatctgacgc atatcgacgc
     2761 agaagtggaa ggcgacaccc atttcccgga ttacgagccg gatgactggg aatcggtatt
     2821 cagcgaattc cacgatgctg atgcgcagaa ctctcacagc tattgctttg agattctgga
     2881 gcggcgggaa caaaaactca tctcagaaga ggatctgtag actagagggc ccgtttaaac
     2941 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3001 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3061 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3121 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3181 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3301 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3421 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3541 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3601 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3661 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3721 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3901 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4021 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     4081 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     4141 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     4201 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     4261 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     4321 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     4381 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     4441 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4501 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4561 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4741 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
