LOCUS       sequence-446050-        9974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of KIF 13a S1371A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9974)
  AUTHORS   Mendoza CS, Plowinske CR, Montgomery AC, Quinones GB, Banker G, 
            Bentley M
  TITLE     Kinesin Regulation in the Proximal Axon is Essential for 
            Dendrite-selective Transport.
  JOURNAL   Mol Biol Cell. 2024 Jun 1;35(6):ar81. doi: 10.1091/mbc.E23-11-0457. 
            Epub 2024 Apr 10.
  PUBMED    38598291
REFERENCE   2  (bases 1 to 9974)
  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: "Mol Biol 
            Cell."; date: "2024-06-1"; volume: "35(6):ar81. doi"; pages: " 
            10.1091/mbc.E23-11-0457. Epub 2024 Apr 10"
COMMENT     Sequence Label: pBa.FRB-3myc-KIF13AS1371A tail 361-1749
FEATURES             Location/Qualifiers
     source          1..9974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..33)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     68..87
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4410..4466
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(4580..4858)
                     /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"
     primer_bind     complement(4938..4960)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     complement(7317..7334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7317..7333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7326..7348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7542..7561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7661..7683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7721..7739)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7807..7911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7912..7980,7981..8772)
                     /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     complement(8130..8149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8943..9531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9432..9451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9685..9702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9819..9840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9855..9860,9861..9878,9879..9885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9893..9909
                     /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)."
     primer_bind     9898..9920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9917..9933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9917..9933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    join(9964..9974,1..124)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
       61 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      121 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      181 tttttaaagc aagtaaaacc tctacagatg tgatatggct gattatgatc attacttatc
      241 tagcttaccg agtggctgtg gcttcctggt cgttctctat cttgggatgc agccggtcgg
      301 catctggagg tctttctggg atacctttat ggtctgtttc gtttgttaga gccctcctcc
      361 tggagggctc ccctacagaa caatctgcca aaccatcaaa gtctttgcca tcagaaacac
      421 ccatgaactc tgtgaaggaa tggtcctcca aaatgtttgt agtgtgttct atcgtcacct
      481 ctcctgggca gaattctctg gcttggtggc catctttgct ggggccgaca tccagttctg
      541 aacatgaagc agtcctgcat ggcattctgg aactttcctc agggatgctc tggggtgacg
      601 gggtcccctt gggtaaggca ctgttttcct ccagcggcac agcgattgtc tcatccttcc
      661 ctgagcctct tcctacttct gtaagctcct gattcgaagc tcgtctgaca tccggtgcaa
      721 gtgatggtcc cgggggctcg gcgcactcga cctcctttgt cgagacggca agctggtctg
      781 agctgtcgct ggaggggact gccatgtcag acagagtggc attggaggca ctgtgggaga
      841 agtagccact ggtgaggctg ctggtggtag ggctacggga cactgctttc tccaaggctc
      901 tcgagtctgt caggtccgcc ttagaagagg accactccct gttctccaag ctggcattgt
      961 aaacactgaa gtcagcgaac tccacgggca cataaggctg tgtaagcatc agcttcctgc
     1021 tgagagcttc cagctcgttt tcctcctctt ccgagtcctc ttggcttagg agaggtctgg
     1081 cttccagggc caaccttttc ttatgctcct cttttacagg catgaggggt ttgaagaact
     1141 ttggtggctg aggagagaac gctttaaaag ggctgactgt taaggcatga ggattctctg
     1201 atggacaacc ttccttgctc ctgcgaggtg agtccctggg taaagtccca tagcacgcga
     1261 cgtcttggta gctggagctg tagtcagaca cgtctaactg gttctcgggc cagcccttat
     1321 cgtcttcatc gaagccagag agatctggcc ggctagagga gacgttgtga acattcggtg
     1381 tcgcgaggct tctccgaatg tgcctcgctt tggtggaaag cgcttccttc accgtgacag
     1441 cctgccggag ccgctccagg ctcaggatat tctccacctg cagcacgccc cgcgtgtact
     1501 tctcgatgta tgtctcccca tccaaagtgc cctcgttctc actccgcgct gccaggaggg
     1561 ccagggtttc ccggtcctca atctcctcgg tcgcctttgg gatattggat actatctcat
     1621 aggtcacgcc gcaggagtaa cgtatattga tcagtgatat tcgtctcttc aaactctggg
     1681 tgaaactctg cttgttataa atattggccg cgatccgctt ccgcaacact agctccatgg
     1741 cggccgggtg gctgagctgc acggtggtct tcacgatcag gtagatcctt tcattctgcg
     1801 gtgtgacccg attcaggtga agagagtcgt gcacagagga gtcccaggag gctgtggctg
     1861 aaacctcgtc atcactgtgc tttatgatgg gcaggtagaa aaactgactg ccgtgttcct
     1921 tgggcaggat ggagttcaca ccggacgcat gggggcccac cagctgctca ttggcactga
     1981 ggtcatccgc gttcagatcc aggaagagaa ccgggatgtg tgtttccatg ccaggaggtg
     2041 ggacccagtc agcaggtgcc ccggggattc cgctgccagg ggcgggcacc agcacagcat
     2101 tcctctcctc tgtcagcccc acccactgct ccaccagcct ggcttcccgc tccatgtcgt
     2161 cttctgtttt ctctttttta ttgctgactt ttttaatctg ttcgtctaag tattcccgtc
     2221 gtttaatgag tgcatcggac catctctctc ttacacaatt caagtcttct tcctggtagc
     2281 tgtccagccc tctctgcagc ttggtggacc tggcggtcac gcagccaatg gatactgata
     2341 ggatggcttc aaccatgagt ggcagtgtcc ctgagtgctg tacaggtttc actgtgacct
     2401 gcactctacg ggagtgaccc tgtctgagtt ggaagacgcc gccggtgttg acgtctttcg
     2461 cctggtgaag ctccacagcg gcataatccc cgagctcatt cagctccaga atggagatcc
     2521 acacctctat tctccgggtc acttcattcc acctgtcatg caatgtcctt gtcttcgcat
     2581 ggagcgagtc cacctcccag atggggcttc cgttcccagc acacctgtgg ccccacactt
     2641 caatcgccag cgcaccatca gaaatgaact ccaggaactc ttctgtgaca gtcaccacgt
     2701 agtccttaca gtgagagaag gtcaccgtgt actgtgcgtc cttggactgc ggggagggca
     2761 catctgggtc caccactgga gcagccactg tagactcaca ctggtcccag aacgtgtact
     2821 gacagaagac gaaatttgag aggcttatgg gtagcccggt tgcctcttta attatcaccc
     2881 tgcatgtcag ctttttgacc cggtgaatga cttcgccgct gctgtcgacc acttcaagac
     2941 tcccgctctc actagagttc tctgaagagt catcctccgc catgcgttct ggaatggttc
     3001 ctgtaatgcg cgtcacctcc acatggaggc gcccggcaac ctccccctgc tggctgatga
     3061 tgggtactgc atattgaagc ttcacatcac agaagagaca ttccaaaaac acattagcca
     3121 caccaatgag gttgtgattc tcctgcgctt cataaaaagg gtcacctcgc ttcccataga
     3181 gcctcttggc ctcaggaaca ttttctttcc attcttggta aaggtctctc atatcaatta
     3241 gcttattctc cagcttctca atggtccaga cctgggtgcc cttccctttc ctcctggctt
     3301 ggatggcggg ctcgctgacg attgcgcctc tcttcctgtt ggcactgagg ttggccgcag
     3361 ggatctgcag agtcacttgg tagtcagtga gtttgctcat ctcctcggct aggaagttgg
     3421 cttccctcac caaagtgttg gctttcacca gctgctctcg aagcttggcc aagctctgcc
     3481 taaagagttc atccctctcc tcggcccact gggtcacctt ctgctgggct gtctggctgc
     3541 tgtaggccag acggtctgag gcgctgctgg gcggctgcct ctcgggagag agctgctggc
     3601 gaagctgctc cagctcgcgt tcgtacatga gccgctgttc ctccagggcg gtccgcttct
     3661 cttccaggta ctgcttctcc aggacctgaa ccacattttg aactggatca ttgctgttca
     3721 gtgttttcat gatgacctcc atctgtgcga actcgtagtt gtagtctggc tccgaggaag
     3781 cctcgctcgc agcgtctaga tcgtgctccg cggaactggt ttctctctca aagtctttca
     3841 accagtcccg ccgtttcctc ttaggcaagt taattctaaa aaagtggttg tttccccata
     3901 agatcctgtc gccgtgccac agctgggtag tactgcagac aagggtgcca ttgccacagg
     3961 accttgcatt ttctttgggc gtgagagtga tgtctccatc cgctgcgatg tcgatctcac
     4021 agtgctcggg ctgaatccct attccgaaga gctggatgtc ctgagaggta tctgcaccca
     4081 cccgggtatg atcctttaaa tagtaaacca gaagctcgtt gagggcaggg tctgcgttca
     4141 ggttgacgag gtagcacttg tcgtccccca ccttgatgcc agaggtctcc agggagattc
     4201 ccatgctttc cagctgccgc tgtctctcct gtgcgatcgc ttctgttttc ctcagcttct
     4261 cttcccacgt caccgttagc tcctttatca gcttctcaga ctcttcgagc ttctccttta
     4321 gctcgggggc tttcatggcc tcagcctgcg agagctgctc cctcagtttc tccacttcct
     4381 ctcgaagctc tcggatgacc ttggcattgg gatcccgggc ccgcggtacc gtcgactgca
     4441 gaattcgaag cttgagctcg agatctgagt acttgtacag atcttcttca gaaataagct
     4501 tttgcagatc ctcttcagaa ataagttttt gcaggtcctc ctctgagatc agcttctgca
     4561 ttccgtacgg ataactagtc tttgagattc gtcggaacac atgataatag aggtcccagg
     4621 cttggaggag gtccttgaca ttccctgatt tcatgtactt cctgcaccac tcttgggcct
     4681 ccattaaatc tcgaccatag gcctgattaa aggatgtttc cttcagagtc tgggggcccc
     4741 gttccatcat agcatgcaag ggctccagca cctcaaacat gcctttcacg ttcctttccc
     4801 caaagtacaa acgagatgcc tcttccaggc cttcatgcca catctcatgc cagaggattc
     4861 tagaagccat ggtaggccta ggcgcgccaa gcttgggggc tgcggaggaa cagagaaggg
     4921 aagatataaa ccccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc
     4981 cgaaggcagc cgtccccctg cggacagccc cgaggctgga gatggagaag gggacggcgg
     5041 cgcggcgacg cacgaaggcc ctccccgccc atttccttcc tgccggcgcc gcaccgcttc
     5101 gccccgcgcc cgctagaggg ggtgcggcgg cgcctcccag atttcggctc cgcacagatt
     5161 tgggacaaag gaagtccctg cgccctctcg cacgattacc ataaaaggca atggctgcgg
     5221 ctcgccgcgc ctcgacagcc gccggcgctc cgggggccgc cgcgcccctc cccccggccc
     5281 ccccccgccc gcggccgccc ccgccccccc cccccccccc ccccccgccc cccccccccc
     5341 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
     5401 cccccccccc gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg
     5461 gggtggggag gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaggtg
     5521 agggtgtggg gcgggggtgt gacgcgcccc tccatccccc ctcccccctt tttgattcca
     5581 cagtgctttg ttggggaggg ggagcggggt ccgcgtgtgc agccctcccc ccacccccga
     5641 ggtggtcctg cggtcccctt gaagtcacct cccggggagc gccagcagga gaggcaggga
     5701 tagcgatggg caatgagcgt gcatgcactg agctgcagcg acactgatgt gcatgcacgc
     5761 accgagcagc aataacaccg gtgtgcgtgc agccactgag ctgcagtgat acaatgagtg
     5821 tgcgtgcacc gagcagcaat cgctccagtg tgagtgcatg cactgggatg aagtgataca
     5881 atgagcgtgc gtgcacagag cagcaataac accgaggggt atgcatgcac tagactgcta
     5941 cacaccaagg aggatgcatg cacagcgctg caccatcaag acgcatgcat gcaccaggct
     6001 gcaataacac caggagcatg catgcacggg cctggtgctg gggtgcagcc tgctgggatg
     6061 caagggaggg gaaggggcac cgggtgcaac gcgcgctgca ctctctacac ccaccccgcc
     6121 cggcctgcag cactacccag ggccaagttc aatgctaatg tcagcacgta gcggtgcaaa
     6181 cctccaggcc ggcccttgga ctgctgagtg cactgctggg tgcacctcag agcccgttaa
     6241 gggccactct ttgtcagggt gtgcgctgcg gcggtggcag ccggagggag gaagcgagcg
     6301 ctgagtaagg agctgagtca ggcagggccc ggcccggcgg aggtcaaggc agccgaatgc
     6361 ctttgcctac atttgggcaa tattttgttt ctttcctgtt tgcagcgcgt gctgcaggct
     6421 gggccggggc ggtgcgcggt gttggtgagc agcgctgcga ggggctttgc agctccagag
     6481 cggtgcatga gcgcatgctg attaactgca ttgattaagg ggccatgcaa gaagcactgt
     6541 agatattagg ggagcgggac ccagccctgt gtgcggtctt gcagtggggt ctttgtccct
     6601 gggatgctgc gaggatgggt tgcatcgggg ctgcattgtg tcccctccaa actggaggtg
     6661 gccctgtgga cagagctgag ctgcagggac accgcaggga caatactgtc tcctcggagg
     6721 cggtgacact gcagcacatg gggacctccg tgggcaccca gtgccaggac cccgtgggct
     6781 ccatagtgcc acatctctct gtatcagccc cactcgcagc aaaaagggag tcacactgtt
     6841 aattacctgc ctgctgggga attaattaac gtttgtgctc cactttgaca aggccgggta
     6901 ttaattagaa gattatacgt caaagaaggg cagccacgag gcatgggtac agcgggaggt
     6961 gtgcagagct gataatgggg cggcaatggg aaatgccccg cggtagccat gaagccgtgg
     7021 gtttgggctg gggaatgtca ccagaaggca gcgtggggtg tgagggcccg gcaggatggg
     7081 ggtggtgggg gcaatgatgg ggcaattcgg gttcccttga ttatggggtt ggatcagagc
     7141 gcggcatccg gagcgtggca gtgccaaccc cactgtgtgc tgctggtagg gggctcctca
     7201 cagtgccact gccatgccac agagccagga ccatcagttt ccaaggggaa ccgggtgctg
     7261 aaggggagca tccccatcct gagagccgtg ggccaatgca aagatcccaa cttggcactg
     7321 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     7381 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     7441 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     7501 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     7561 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7621 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7681 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     7741 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7801 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7861 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7921 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7981 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8041 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8101 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8161 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8221 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8281 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8341 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8401 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8461 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8521 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8581 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8641 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8701 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8761 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8821 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8881 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9061 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     9601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     9661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     9721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     9781 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     9841 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     9901 ggataacaat ttcacacagg aaacagctat gacatgatta cgaattcgag ctcggtaccc
     9961 ggggatccag acat
//
