LOCUS       sequence-380191-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3A CC 419-589.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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-KIF3A CC 419-589
FEATURES             Location/Qualifiers
     source          1..7284
                     /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    2703..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2740..2759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2794..2813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2868..2884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2881..2903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2892..2908
                     /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(2916..2922,2923..2940,2941..2946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2961..2982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3099..3116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3270..3858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3350..3369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4029..4820,4821..4889))
                     /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     4652..4671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4890..4994)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5062..5080
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5118..5140)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5240..5259
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5453..5475
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5467..5484
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5468..5484
                     /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 caagccagac aagatgattg aaatgcaagc aaaaattgat
     2101 gaggagagaa aagcacttga aacaaagctc gacatggaag aagaagaaag aaacaaggct
     2161 agagctgaat tagagaaacg ggaaaaagat cttcttaaag cccaacaaga gcatcagtct
     2221 ttgctggaaa aattatctgc cctggaaaag aaggtaattg ttggtggggt tgacttgttg
     2281 gccaaagctg aggaacaaga gaaacttctt gaagaatcta acatggaact ggaagaaagg
     2341 aggaaaaggg cagagcaact tcgcagagaa cttgaggaaa aagagcaaga acgcttggat
     2401 attgaagaaa aatataccag tttgcaagag gaagcacagg gaaagaccaa gaagttaaag
     2461 aaagtttgga ctatgctgat ggctgcaaag tcagagatgg ctgatctcca acaagaacat
     2521 cagagggaaa ttgaaggcct actggagaac attcggcaac ttagccggga gctttaattt
     2581 aatctagata agtaatgatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     2641 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2701 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2761 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2821 cttatcatgt ctggatcccc gggtaccgag ctcgaattcg taatcatgtc atagctgttt
     2881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4021 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4081 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4141 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4201 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4261 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4321 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4381 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4441 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4501 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4561 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4621 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4681 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4741 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4801 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4861 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4921 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4981 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5041 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5101 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5161 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5221 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5281 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5341 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5401 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5461 cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc cacggctctc
     5521 aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg tcctggctct
     5581 gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg gggttggcac
     5641 tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc
     5701 ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttctgg
     5761 tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg
     5821 ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg
     5881 acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat
     5941 tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac
     6001 agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc
     6061 catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt gtccctgcag
     6121 ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca
     6181 acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg
     6241 gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa tgcagttaat
     6301 cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct gctcaccaac
     6361 accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag aaacaaaata
     6421 ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc gggccctgcc
     6481 tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc cgcagcgcac
     6541 accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg cactcagcag
     6601 tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt gaacttggcc
     6661 ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc gttgcacccg
     6721 gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg cccgtgcatg
     6781 catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg cagcgctgtg
     6841 catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt gttattgctg
     6901 ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca cactggagcg
     6961 attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct gcacgcacac
     7021 cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct cagtgcatgc
     7081 acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg aggtgacttc
     7141 aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg accccgctcc
     7201 ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga ggggcgcgtc
     7261 acacccccgc cccacaccct cacc
//
