LOCUS       sequence-380427-        7385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3C CC 411-615.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Garbouchian A, Montgomery A, Gilbert SP, Bentley M
  TITLE     KAP is the neuronal organelle adaptor for Kinesin-2 KIF3AB and 
            KIF3AC.
  JOURNAL   Mol Biol Cell. 2022 Oct 6:mbcE22080336. doi: 
            10.1091/mbc.E22-08-0336.
  PUBMED    36200888
REFERENCE   2  (bases 1 to 7385)
  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. 2022 Oct 6:mbcE22080336. doi: 10.1091/mbc.E22-08-0336."
COMMENT     Sequence Label: pBa.GFP-KIF3C CC 411-615
FEATURES             Location/Qualifiers
     source          1..7385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..279
                     /label=chicken beta-actin promoter
     primer_bind     1222..1244
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             join(1312..1314,1315..1317,1318..2025)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGGELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1357..1378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1618..1637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1965..1986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2804..2938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2841..2860)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2895..2914
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2969..2985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2969..2985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2982..3004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2993..3009
                     /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(3017..3023,3024..3041,3042..3047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3062..3083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3200..3217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3371..3959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3451..3470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4130..4921,4922..4990))
                     /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     4753..4772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4991..5095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5163..5181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5219..5241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5341..5360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5554..5576
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5568..5585
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5569..5585
                     /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 gggggggggg
      121 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
      181 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      241 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      301 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      361 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      421 gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg gctccgggag
      481 ggccctttgt gcggggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      541 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
      601 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
      661 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
      721 gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc cgagttgctg
      781 agcacggccc ggcttcgggt gcggggctcc gtgcggggcg tggcgcgggg ctcgccgtgc
      841 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
      901 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
      961 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1021 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1081 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1141 gccgccgtcc ccttctccat ctccagcctc ggggctgtcc gcagggggac ggctgccttc
     1201 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggg gtttatatct
     1261 tcccttctct gttcctccgc agcccccaag cttggcgcgc ctaggcctac catggtgagc
     1321 aagggcgggg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1381 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1441 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1501 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1561 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1621 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1681 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1741 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1801 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1861 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1921 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1981 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacggcgg aggatccgga
     2041 ggcggctctg gcggagggct gtacagtcgt agaaagaagg ccgtgtctgc cccagccggc
     2101 tatccagagg ggtccgtgat tgaggcctgg gtggccgaag aggaagatga taacaacaac
     2161 aatcaccacc caccccagcc catcttggag gcagctttgg agaagaacat ggagaattac
     2221 ctgcaggacc agaaggaacg gctggaggag gagaaggctg ccatccagga tgaccgcagc
     2281 cttgtgagtg aggagaagca gaagcttcta gaagagaagg agaagatgct ggaggatcta
     2341 cggcgggagc agcaggctac agagctgctt gcagccaagt acaaggccat ggaaagcaag
     2401 ctactcattg gggggcggaa catcatggat cacaccaacg agcaacagaa gatgttggaa
     2461 ctgaagcgac aggagattgc ggaacagaaa cgccgtgaac gggaaatgca gcaggagatg
     2521 ctgctccgcg atgaagaaac catggagcta cgtggtacct actcatccct gcagcaggag
     2581 gtggaagtta aaaccaagaa actcaagaag ctctatgcca agctacaggc ggtgaaggcc
     2641 gagatccagg accaacacga agagtacatc cgcgtgcgtc taatctagat aagtaatgat
     2701 cataatcagc catatcacat ctgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2761 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccc
     2941 cgggtaccga gctcgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     3001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3181 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3241 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3301 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3541 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3781 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5101 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5161 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5221 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5281 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5341 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5401 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     5461 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5521 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5581 ccagtgccaa gttgggatct ttgcattggc ccacggctct caggatgggg atgctcccct
     5641 tcagcacccg gttccccttg gaaactgatg gtcctggctc tgtggcatgg cagtggcact
     5701 gtgaggagcc ccctaccagc agcacacagt ggggttggca ctgccacgct ccggatgccg
     5761 cgctctgatc caaccccata atcaagggaa cccgaattgc cccatcattg cccccaccac
     5821 ccccatcctg ccgggccctc acaccccacg ctgccttctg gtgacattcc ccagcccaaa
     5881 cccacggctt catggctacc gcggggcatt tcccattgcc gccccattat cagctctgca
     5941 cacctcccgc tgtacccatg cctcgtggct gcccttcttt gacgtataat cttctaatta
     6001 atacccggcc ttgtcaaagt ggagcacaaa cgttaattaa ttccccagca ggcaggtaat
     6061 taacagtgtg actccctttt tgctgcgagt ggggctgata cagagagatg tggcactatg
     6121 gagcccacgg ggtcctggca ctgggtgccc acggaggtcc ccatgtgctg cagtgtcacc
     6181 gcctccgagg agacagtatt gtccctgcgg tgtccctgca gctcagctct gtccacaggg
     6241 ccacctccag tttggagggg acacaatgca gccccgatgc aacccatcct cgcagcatcc
     6301 cagggacaaa gaccccactg caagaccgca cacagggctg ggtcccgctc ccctaatatc
     6361 tacagtgctt cttgcatggc cccttaatca atgcagttaa tcagcatgcg ctcatgcacc
     6421 gctctggagc tgcaaagccc ctcgcagcgc tgctcaccaa caccgcgcac cgccccggcc
     6481 cagcctgcag cacgcgctgc aaacaggaaa gaaacaaaat attgcccaaa tgtaggcaaa
     6541 ggcattcggc tgccttgacc tccgccgggc cgggccctgc ctgactcagc tccttactca
     6601 gcgctcgctt cctccctccg gctgccaccg ccgcagcgca caccctgaca aagagtggcc
     6661 cttaacgggc tctgaggtgc acccagcagt gcactcagca gtccaagggc cggcctggag
     6721 gtttgcaccg ctacgtgctg acattagcat tgaacttggc cctgggtagt gctgcaggcc
     6781 gggcggggtg ggtgtagaga gtgcagcgcg cgttgcaccc ggtgcccctt cccctccctt
     6841 gcatcccagc aggctgcacc ccagcaccag gcccgtgcat gcatgctcct ggtgttattg
     6901 cagcctggtg catgcatgcg tcttgatggt gcagcgctgt gcatgcatcc tccttggtgt
     6961 gtagcagtct agtgcatgca tacccctcgg tgttattgct gctctgtgca cgcacgctca
     7021 ttgtatcact tcatcccagt gcatgcactc acactggagc gattgctgct cggtgcacgc
     7081 acactcattg tatcactgca gctcagtggc tgcacgcaca ccggtgttat tgctgctcgg
     7141 tgcgtgcatg cacatcagtg tcgctgcagc tcagtgcatg cacgctcatt gcccatcgct
     7201 atccctgcct ctcctgctgg cgctccccgg gaggtgactt caaggggacc gcaggaccac
     7261 ctcgggggtg gggggagggc tgcacacgcg gaccccgctc cccctcccca acaaagcact
     7321 gtggaatcaa aaagggggga ggggggatgg aggggcgcgt cacacccccg ccccacaccc
     7381 tcacc
//
