LOCUS       sequence-380540-        7327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3C DD 616-796.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7327)
  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 7327)
  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 DD 616-796
FEATURES             Location/Qualifiers
     source          1..7327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..276
                     /label=chicken beta-actin promoter
     primer_bind     1219..1241
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             join(1309..1311,1312..1314,1315..2022)
                     /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(1354..1375)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1615..1634)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1962..1983
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2746..2880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2837..2856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2911..2927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2911..2927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2924..2946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2935..2951
                     /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(2959..2965,2966..2983,2984..2989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3004..3025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3142..3159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3313..3901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3393..3412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4072..4863,4864..4932))
                     /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     4695..4714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4933..5037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5105..5123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5161..5183)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5283..5302
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5496..5518
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5510..5527
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5511..5527
                     /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 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      181 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      241 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      301 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      361 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      421 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
      481 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      541 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
      601 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
      661 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
      721 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
      781 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg
      841 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
      901 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
      961 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1021 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1081 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1141 gccgtcccct tctccatctc cagcctcggg gctgtccgca gggggacggc tgccttcggg
     1201 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggggtt tatatcttcc
     1261 cttctctgtt cctccgcagc ccccaagctt ggcgcgccta ggcctaccat ggtgagcaag
     1321 ggcggggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1381 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1441 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1501 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1561 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1621 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1681 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1741 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1801 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1861 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1921 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1981 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acggcggagg atccggaggc
     2041 ggctctggcg gagggctgta cactcggcag gacctggagg aggcgcagaa tgagcagacc
     2101 cgggaactaa agctcaagta cctcatcatt gagaacttca tcccccctga ggagaagaac
     2161 aagatcatga accgcctttt cctggactgt gaggaggagc agtggaggtt ccagccactg
     2221 gtgccagctg gagtgaataa cagccaaatg aagaagcgtc caacttctgc ggtgggctac
     2281 aagagaccta tcagccagta tgctcgggtt gccatggcaa tggggtccca tcccagatac
     2341 agagctgaaa atataatgtt tttggaactg gatgtgtccc caccggctat ctttgagatg
     2401 gagttttctc atgaccaaga gcaagaccca cgggtactac acatggagag gctcatgagg
     2461 ttggacagct ttctggaaag accgtccaca actaaagtgc gaaagtcgag atcctggtgc
     2521 cagagtcctc agcggatgcc tccaccctcc acagctcatg catccatgac ctctgttcct
     2581 ttgcaccctg caacagtggt agaccatgac tgacggaccg tctaatctag ataagtaatg
     2641 atcataatca gccatatcac atctgtagag gttttacttg ctttaaaaaa cctcccacac
     2701 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     2881 cccgggtacc gagctcgaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     2941 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3001 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3061 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3121 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3181 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3301 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3361 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3421 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3481 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3541 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3601 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3661 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3721 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3781 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3841 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3961 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5041 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5101 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5161 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5221 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5281 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5341 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5401 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5461 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5521 ggccagtgcc aagttgggat ctttgcattg gcccacggct ctcaggatgg ggatgctccc
     5581 cttcagcacc cggttcccct tggaaactga tggtcctggc tctgtggcat ggcagtggca
     5641 ctgtgaggag ccccctacca gcagcacaca gtggggttgg cactgccacg ctccggatgc
     5701 cgcgctctga tccaacccca taatcaaggg aacccgaatt gccccatcat tgcccccacc
     5761 acccccatcc tgccgggccc tcacacccca cgctgccttc tggtgacatt ccccagccca
     5821 aacccacggc ttcatggcta ccgcggggca tttcccattg ccgccccatt atcagctctg
     5881 cacacctccc gctgtaccca tgcctcgtgg ctgcccttct ttgacgtata atcttctaat
     5941 taatacccgg ccttgtcaaa gtggagcaca aacgttaatt aattccccag caggcaggta
     6001 attaacagtg tgactccctt tttgctgcga gtggggctga tacagagaga tgtggcacta
     6061 tggagcccac ggggtcctgg cactgggtgc ccacggaggt ccccatgtgc tgcagtgtca
     6121 ccgcctccga ggagacagta ttgtccctgc ggtgtccctg cagctcagct ctgtccacag
     6181 ggccacctcc agtttggagg ggacacaatg cagccccgat gcaacccatc ctcgcagcat
     6241 cccagggaca aagaccccac tgcaagaccg cacacagggc tgggtcccgc tcccctaata
     6301 tctacagtgc ttcttgcatg gccccttaat caatgcagtt aatcagcatg cgctcatgca
     6361 ccgctctgga gctgcaaagc ccctcgcagc gctgctcacc aacaccgcgc accgccccgg
     6421 cccagcctgc agcacgcgct gcaaacagga aagaaacaaa atattgccca aatgtaggca
     6481 aaggcattcg gctgccttga cctccgccgg gccgggccct gcctgactca gctccttact
     6541 cagcgctcgc ttcctccctc cggctgccac cgccgcagcg cacaccctga caaagagtgg
     6601 cccttaacgg gctctgaggt gcacccagca gtgcactcag cagtccaagg gccggcctgg
     6661 aggtttgcac cgctacgtgc tgacattagc attgaacttg gccctgggta gtgctgcagg
     6721 ccgggcgggg tgggtgtaga gagtgcagcg cgcgttgcac ccggtgcccc ttcccctccc
     6781 ttgcatccca gcaggctgca ccccagcacc aggcccgtgc atgcatgctc ctggtgttat
     6841 tgcagcctgg tgcatgcatg cgtcttgatg gtgcagcgct gtgcatgcat cctccttggt
     6901 gtgtagcagt ctagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct
     6961 cattgtatca cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac
     7021 gcacactcat tgtatcactg cagctcagtg gctgcacgca caccggtgtt attgctgctc
     7081 ggtgcgtgca tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg
     7141 ctatccctgc ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc
     7201 acctcggggg tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca
     7261 ctgtggaatc aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac
     7321 cctcacc
//
