LOCUS       sequence-446051-        9738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of KIF 13a WLDLE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9738)
  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 9738)
  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-KIF13AWLDLE tail 361-1749
FEATURES             Location/Qualifiers
     source          1..9738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1799..1816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1799..1815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2024..2043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2143..2165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2203..2221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2289..2393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2394..2462,2463..3254)
                     /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(2612..2631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3425..4013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3914..3933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4167..4184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4301..4322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4337..4342,4343..4360,4361..4367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4375..4391
                     /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     4380..4402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4399..4415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4399..4415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4446..4580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4470..4489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4524..4543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      6188..6197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    8863..8919
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(9033..9311)
                     /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(9391..9413)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 tgagggtgtg gggcgggggt gtgacgcgcc cctccatccc ccctcccccc tttttgattc
       61 cacagtgctt tgttggggag ggggagcggg gtccgcgtgt gcagccctcc ccccaccccc
      121 gaggtggtcc tgcggtcccc ttgaagtcac ctcccgggga gcgccagcag gagaggcagg
      181 gatagcgatg ggcaatgagc gtgcatgcac tgagctgcag cgacactgat gtgcatgcac
      241 gcaccgagca gcaataacac cggtgtgcgt gcagccactg agctgcagtg atacaatgag
      301 tgtgcgtgca ccgagcagca atcgctccag tgtgagtgca tgcactggga tgaagtgata
      361 caatgagcgt gcgtgcacag agcagcaata acaccgaggg gtatgcatgc actagactgc
      421 tacacaccaa ggaggatgca tgcacagcgc tgcaccatca agacgcatgc atgcaccagg
      481 ctgcaataac accaggagca tgcatgcacg ggcctggtgc tggggtgcag cctgctggga
      541 tgcaagggag gggaaggggc accgggtgca acgcgcgctg cactctctac acccaccccg
      601 cccggcctgc agcactaccc agggccaagt tcaatgctaa tgtcagcacg tagcggtgca
      661 aacctccagg ccggcccttg gactgctgag tgcactgctg ggtgcacctc agagcccgtt
      721 aagggccact ctttgtcagg gtgtgcgctg cggcggtggc agccggaggg aggaagcgag
      781 cgctgagtaa ggagctgagt caggcagggc ccggcccggc ggaggtcaag gcagccgaat
      841 gcctttgcct acatttgggc aatattttgt ttctttcctg tttgcagcgc gtgctgcagg
      901 ctgggccggg gcggtgcgcg gtgttggtga gcagcgctgc gaggggcttt gcagctccag
      961 agcggtgcat gagcgcatgc tgattaactg cattgattaa ggggccatgc aagaagcact
     1021 gtagatatta ggggagcggg acccagccct gtgtgcggtc ttgcagtggg gtctttgtcc
     1081 ctgggatgct gcgaggatgg gttgcatcgg ggctgcattg tgtcccctcc aaactggagg
     1141 tggccctgtg gacagagctg agctgcaggg acaccgcagg gacaatactg tctcctcgga
     1201 ggcggtgaca ctgcagcaca tggggacctc cgtgggcacc cagtgccagg accccgtggg
     1261 ctccatagtg ccacatctct ctgtatcagc cccactcgca gcaaaaaggg agtcacactg
     1321 ttaattacct gcctgctggg gaattaatta acgtttgtgc tccactttga caaggccggg
     1381 tattaattag aagattatac gtcaaagaag ggcagccacg aggcatgggt acagcgggag
     1441 gtgtgcagag ctgataatgg ggcggcaatg ggaaatgccc cgcggtagcc atgaagccgt
     1501 gggtttgggc tggggaatgt caccagaagg cagcgtgggg tgtgagggcc cggcaggatg
     1561 ggggtggtgg gggcaatgat ggggcaattc gggttccctt gattatgggg ttggatcaga
     1621 gcgcggcatc cggagcgtgg cagtgccaac cccactgtgt gctgctggta gggggctcct
     1681 cacagtgcca ctgccatgcc acagagccag gaccatcagt ttccaagggg aaccgggtgc
     1741 tgaaggggag catccccatc ctgagagccg tgggccaatg caaagatccc aacttggcac
     1801 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1861 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1921 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     1981 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     2041 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2101 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2161 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2221 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4261 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     4321 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     4381 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattcg agctcggtac
     4441 ccggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4501 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4561 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4621 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta cagatgtgat atggctgatt
     4681 atgatcatta cttatctagc ttaccgagtg gctgtggctt cctggtcgtt ctctatcttg
     4741 ggatgcagcc ggtcggcatc tggaggtctt tctgggatac ctttatggtc tgtttcgttt
     4801 gttagagccc tcctcctgga gggctcccct acagaacaat ctgccaaacc atcaaagtct
     4861 ttgccatcag aaacacccat gaactctgtg aaggaatggt cctccaaaat gtttgtagtg
     4921 tgttctatcg tcacctctcc tgggcagaat tctctggctt ggtggccatc tttgctgggg
     4981 ccgacatcca gttctgaaca tgaagcagtc ctgcatggca ttctggaact ttcctcaggg
     5041 atgctctggg gtgacggggt ccccttgggt aaggcactgt tttcctccag cggcacagcg
     5101 attgtctcat ccttccctga gcctcttcct acttctgtaa gctcctgatt cgaagctcgt
     5161 ctgacatccg gtgcaagtga tggtcccggg ggctcggcgc actcgacctc ctttgtcgag
     5221 acggcaagct ggtctgagct gtcgctggag gggactgcca tgtcagacag agtggcattg
     5281 gaggcactgt gggagaagta gccactggtg aggctgctgg tggtagggct acgggacact
     5341 gctttctcca aggctctcga gtctgtcagg tccgccttag aagaggacca ctccctgttc
     5401 tccaagctgg cattgtaaac actgaagtca gcgaactcca cgggcacata aggctgtgta
     5461 agcatcagct tcctgctgag agcttccagc tcgttttcct cctcttccga gtcctcttgg
     5521 cttaggagag gtctggcttc cagggccaac cttttcttat gctcctcttt tacaggcatg
     5581 aggggtttga agaactttgg tggctgagga gagaacgctt taaaagggct gactgttaag
     5641 gcatgaggat tctctgatgg acaaccttcc ttgctcctgc gaggtgagtc cctgggtaaa
     5701 gtcccatagc acgcgacgtc ttggtagctg gagctgtagt cagacacgtc taactggttc
     5761 tcgggccagc ccttatcgtc ttcatcgaag ccagagagat ctggccggct agaggagacg
     5821 ttgtgaacat tctcgaggtc gagccaccga atgtgcctcg ctttggtgga aagcgcttcc
     5881 ttcaccgtga cagcctgccg gagccgctcc aggctcagga tattctccac ctgcagcacg
     5941 ccccgcgtgt acttctcgat gtatgtctcc ccatccaaag tgccctcgtt ctcactccgc
     6001 gctgccagga gggccagggt ttcccggtcc tcaatctcct cggtcgcctt tgggatattg
     6061 gatactatct cataggtcac gccgcaggag taacgtatat tgatcagtga tattcgtctc
     6121 ttcaaactct gggtgaaact ctgcttgtta taaatattgg ccgcgatccg cttccgcaac
     6181 actagctcca tggcggccgg gtggctgagc tgcacggtgg tcttcacgat caggtagatc
     6241 ctttcattct gcggtgtgac ccgattcagg tgaagagagt cgtgcacaga ggagtcccag
     6301 gaggctgtgg ctgaaacctc gtcatcactg tgctttatga tgggcaggta gaaaaactga
     6361 ctgccgtgtt ccttgggcag gatggagttc acaccggacg catgggggcc caccagctgc
     6421 tcattggcac tgaggtcatc cgcgttcaga tccaggaaga gaaccgggat gtgtgtttcc
     6481 atgccaggag gtgggaccca gtcagcaggt gccccgggga ttccgctgcc aggggcgggc
     6541 accagcacag cattcctctc ctctgtcagc cccacccact gctccaccag cctggcttcc
     6601 cgctccatgt cgtcttctgt tttctctttt ttattgctga cttttttaat ctgttcgtct
     6661 aagtattccc gtcgtttaat gagtgcatcg gaccatctct ctcttacaca attcaagtct
     6721 tcttcctggt agctgtccag ccctctctgc agcttggtgg acctggcggt cacgcagcca
     6781 atggatactg ataggatggc ttcaaccatg agtggcagtg tccctgagtg ctgtacaggt
     6841 ttcactgtga cctgcactct acgggagtga ccctgtctga gttggaagac gccgccggtg
     6901 ttgacgtctt tcgcctggtg aagctccaca gcggcataat ccccgagctc attcagctcc
     6961 agaatggaga tccacacctc tattctccgg ttcacttcat tccacctgtc atgcaatgtc
     7021 cttgtcttcg catggagcga gtccacctcc cagatggggc ttccgttccc agcacacctg
     7081 tggccccaca cttcaatcgc cagcgcacca tcagaaatga actccaggaa ctcttctgtg
     7141 acagtcacca cgtagtcctt acagtgagag aaggtcaccg tgtactgtgc gtccttggac
     7201 tgcggggagg gcacatctgg gtccaccact ggagcagcca ctgtagactc acactggtcc
     7261 cagaacgtgt actgacagaa gacgaaattt gagaggctta tgggtagccc ggttgcctct
     7321 ttaattatca ccctgcatgt cagctttttg acccggtgaa tgacttcgcc gctgctgtcg
     7381 accacttcaa gactcccgct ctcactagag ttctctgaag agtcatcctc cgccatgcgt
     7441 tctggaatgg ttcctgtaat gcgcgtcacc tccacatgga ggcgcccggc aacctccccc
     7501 tgctggctga tgatgggtac tgcatattga agcttcacat cacagaagag acattccaaa
     7561 aacacattag ccacaccaat gaggttgtga ttctcctgcg cttcataaaa agggtcacct
     7621 cgcttcccat agagcctctt ggcctcagga acattttctt tccattcttg gtaaaggtct
     7681 ctcatatcaa ttagcttatt ctccagcttc tcaatggtcc agacctgggt gcccttccct
     7741 ttcctcctgg cttggatggc gggctcgctg acgattgcgc ctctcttcct gttggcactg
     7801 aggttggccg cagggatctg cagagtcact tggtagtcag tgagtttgct catctcctcg
     7861 gctaggaagt tggcttccct caccaaagtg ttggctttca ccagctgctc tcgaagcttg
     7921 gccaagctct gcctaaagag ttcatccctc tcctcggccc actgggtcac cttctgctgg
     7981 gctgtctggc tgctgtaggc cagacggtct gaggcgctgc tgggcggctg cctctcggga
     8041 gagagctgct ggcgaagctg ctccagctcg ccttcgtaca tgagccgctg ttcctccagg
     8101 gcggtccgct tctcttccag gtactgcttc tccaggacct gaaccacatt ttgaactgga
     8161 tcattgctgt tcagtgtttt catgatgacc tccatctgtg cgaactcgta gttgtagtct
     8221 ggctccgagg aagcctcgct cgcagcgtct agatcgtgct ccgcggaact ggtttctctc
     8281 tcaaagtctt tcaaccagtc ccgccgtttc ctcttaggca agttaattct aaaaaagtgg
     8341 ttgtttcccc ataagatcct gtcgccgtgc cacagctggg tagtactgca gacaagggtg
     8401 ccattgccac aggaccttgc attttctttg ggcgtgagag tgatgtctcc atccgctgcg
     8461 atgtcgatct cacagtgctc gggctgaatc cctattccga agagctggat gtcctgagag
     8521 gtatctgcac ccacccgggt atgatccttt aaatagtaaa ccagaagctc gttgagggca
     8581 gggtctgcgt tcaggttgac gaggtagcac ttgtcgtccc ccaccttgat gccagaggtc
     8641 tccagggaga ttcccatgct ttccagctgc cgctgtctct cctgtgcgat cgcttctgtt
     8701 ttcctcagct tctcttccca cgtcaccgtt agctccttta tcagcttctc agactcttcg
     8761 agcttctcct ttagctcggg ggctttcatg gcctcagcct gcgagagctg ctccctcagt
     8821 ttctccactt cctctcgaag ctctcggatg accttggcat tgggatcccg ggcccgcggt
     8881 accgtcgact gcagaattcg aagcttgagc tcgagatctg agtacttgta cagatcttct
     8941 tcagaaataa gcttttgcag atcctcttca gaaataagtt tttgcaggtc ctcctctgag
     9001 atcagcttct gcattccgta cggataacta gtctttgaga ttcgtcggaa cacatgataa
     9061 tagaggtccc aggcttggag gaggtccttg acattccctg atttcatgta cttcctgcac
     9121 cactcttggg cctccattaa atctcgacca taggcctgat taaaggatgt ttccttcaga
     9181 gtctgggggc cccgttccat catagcatgc aagggctcca gcacctcaaa catgcctttc
     9241 acgttccttt ccccaaagta caaacgagat gcctcttcca ggccttcatg ccacatctca
     9301 tgccagagga ttctagaagc catggtaggc ctaggcgcgc caagcttggg ggctgcggag
     9361 gaacagagaa gggaagatat aaaccccgcc ggtcacacgc cagaagccga accccgccct
     9421 gccccgtccc ccccgaaggc agccgtcccc ctgcggacag ccccgaggct ggagatggag
     9481 aaggggacgg cggcgcggcg acgcacgaag gccctccccg cccatttcct tcctgccggc
     9541 gccgcaccgc ttcgccccgc gcccgctaga gggggtgcgg cggcgcctcc cagatttcgg
     9601 ctccgcacag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt accataaaag
     9661 gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccgggggc cgccgcgccc
     9721 cccccccccc cccccccc
//
