LOCUS       sequence-267829-        8064 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of mouse KLC1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8064)
  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 8064)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pB.FRB-3myc-KLC1a
FEATURES             Location/Qualifiers
     source          1..8064
                     /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"
     polyA_signal    3483..3617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3520..3539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3574..3593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3648..3664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3661..3683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3672..3688
                     /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(3696..3702,3703..3720,3721..3726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3741..3762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3879..3896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4050..4638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4130..4149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4809..5600,5601..5669))
                     /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     5432..5451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5670..5774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5842..5860
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5898..5920)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6020..6039
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6233..6255
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6247..6264
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6248..6264
                     /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 gcttctagaa
     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 tacggaatgc
     1621 agaagctgat ctcagaggag gacctgcaaa aacttatttc tgaagaggat ctgcaaaagc
     1681 ttatttctga agaagatctg tacaagggag gatccggagg catgtatgac aacatgtcca
     1741 ccatggtgta cataaaggaa gagaagctgg agaagctcac gcaggatgag atcatctcta
     1801 agaccaagca agtgatccag gggctggaag ccctgaagaa tgagcacaac tccatcctgc
     1861 agagtttgct ggagacgctg aagtgcttga agaaggatga cgagagcaac ctggtggaag
     1921 agaaatccag catgatccgc aagtccctgg agatgctgga gcttggcctg agcgaggcgc
     1981 aggtgatgat ggcgctgtcc aatcacctga atgctgtgga gtccgagaag caaaagctcc
     2041 gcgctcaggt tcgacggctg tgccaggaga accagtggct gcgggatgag ctggccaaca
     2101 cgcagcagaa gttgcagaag agcgagcagt cggtggcgca gctggaggag gagaagaaac
     2161 acctggagtt catgaaccag ctgaagaagt acgacgacga catctccccc tcggaggaca
     2221 aagactctga ttcttccaaa gagccgttgg atgatctctt cccaaatgac gaggacgaac
     2281 caggacaagg aatccagcag cagcacagta gtgctgcggc cgccgcccag cagggcggct
     2341 acgagatccc tgcaaggctg cgcacgctcc acaacttggt gatccagtat gcttcacagg
     2401 ggcgttacga ggtggcggtg ccactctgca agcaggccct ggaggatctg gagaagactt
     2461 ccggccacga ccaccccgat gtggctacca tgctcaacat cttggccctg gtgtacaggg
     2521 atcagaacaa gtataaagat gcagctaacc tcctgaacga cgccctggct atccgcgaga
     2581 aaaccctggg cagagatcac cccgcggtgg cagcgactct gaacaaccta gcagtactgt
     2641 acggtaagcg agggaagtac aaggaggcgg agccgctgtg taaacgagcc ctggagatca
     2701 gggagaaggt tctgggaaag gatcatcctg atgttgccaa acagttaaat aacctggccc
     2761 tgctgtgcca gaaccagggc aagtacgagg aggtggagta ttattaccag agggccctgg
     2821 gcatctacca gacgaagctg gggcccgacg atcccaacgt ggccaagacc aagaacaacc
     2881 tggcctcctg ttatctgaaa caagggaagt tcaagcaggc agaaacgctg tacaaggaga
     2941 ttctcacccg cgcacacgag cgggagtttg gatctgtgga cgacgagaac aagcccatct
     3001 ggatgcacgc tgaagagaga gaggagtgca aaggcaagca gaaggacggg tcggcttttg
     3061 gagagtatgg cggctggtat aaagcctgca aagtggacag tcccaccgtc acaaccacct
     3121 tgaaaaacct tggagcactt taccgacggc aggggaagtt tgaagctgca gagacattgg
     3181 aagaagccgc catgaggtca cgtaagcagg gtcttgacaa tgttcacaaa cagagagtgg
     3241 ctgaagtgct aaatgaccct gagagcatgg agaagcggag gagccgggag agtctcaata
     3301 tggacgtggt caagtacgag agtggccctg acggagggga ggaagcctag cggaccgtct
     3361 aatctagata agtaatgatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     3421 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3481 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3601 cttatcatgt ctggatcccc gggtaccgag ctcgaattcg taatcatgtc atagctgttt
     3661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5101 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5401 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5761 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5821 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5881 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5941 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6001 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6061 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     6121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6181 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6241 cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc cacggctctc
     6301 aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg tcctggctct
     6361 gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg gggttggcac
     6421 tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc
     6481 ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttctgg
     6541 tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg
     6601 ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg
     6661 acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat
     6721 tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac
     6781 agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc
     6841 catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt gtccctgcag
     6901 ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca
     6961 acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg
     7021 gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa tgcagttaat
     7081 cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct gctcaccaac
     7141 accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag aaacaaaata
     7201 ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc gggccctgcc
     7261 tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc cgcagcgcac
     7321 accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg cactcagcag
     7381 tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt gaacttggcc
     7441 ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc gttgcacccg
     7501 gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg cccgtgcatg
     7561 catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg cagcgctgtg
     7621 catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt gttattgctg
     7681 ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca cactggagcg
     7741 attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct gcacgcacac
     7801 cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct cagtgcatgc
     7861 acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg aggtgacttc
     7921 aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg accccgctcc
     7981 ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga ggggcgcgtc
     8041 acacccccgc cccacaccct cacc
//
