LOCUS       sequence-380400-        7123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3A DD 590-702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7123)
  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 7123)
  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 DD 590-702
FEATURES             Location/Qualifiers
     source          1..7123
                     /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    2542..2676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2579..2598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2633..2652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2707..2723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2707..2723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2720..2742)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2731..2747
                     /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(2755..2761,2762..2779,2780..2785))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2938..2955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3109..3697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3189..3208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3868..4659,4660..4728))
                     /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     4491..4510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4729..4833)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4901..4919
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4957..4979)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5079..5098
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5292..5314
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5306..5323
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5307..5323
                     /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 catccgactt cagatgctta ttattgataa ctttatacct
     2101 cgggattatc aggaaatgat tgaaaactat gtccattgga atgaagacat aggagaatgg
     2161 cagctaaaat gtgttgctta tacaggaaat aacatgagga agcaaacccc agtacctgat
     2221 aaaaaggaga aagatccctt tgaggtggac ctttctcacg tgtatcttgc ctatactgag
     2281 gagagtctgc gtcagtcttt gatgaaacta gaaagaccac gaacttcaaa ggggaaagca
     2341 aggccaaaga cagggagaag aaagcgttct gcaaagcctg aaactgtaat tgactctctg
     2401 ctgcagtaac ggaccgtcta atctagataa gtaatgatca taatcagcca tatcacatct
     2461 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2521 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2581 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2641 tccaaactca tcaatgtatc ttatcatgtc tggatccccg ggtaccgagc tcgaattcgt
     2701 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2761 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2821 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2881 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2941 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3301 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3541 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4861 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4921 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4981 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5041 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5101 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5161 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5221 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5281 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagt tgggatcttt
     5341 gcattggccc acggctctca ggatggggat gctccccttc agcacccggt tccccttgga
     5401 aactgatggt cctggctctg tggcatggca gtggcactgt gaggagcccc ctaccagcag
     5461 cacacagtgg ggttggcact gccacgctcc ggatgccgcg ctctgatcca accccataat
     5521 caagggaacc cgaattgccc catcattgcc cccaccaccc ccatcctgcc gggccctcac
     5581 accccacgct gccttctggt gacattcccc agcccaaacc cacggcttca tggctaccgc
     5641 ggggcatttc ccattgccgc cccattatca gctctgcaca cctcccgctg tacccatgcc
     5701 tcgtggctgc ccttctttga cgtataatct tctaattaat acccggcctt gtcaaagtgg
     5761 agcacaaacg ttaattaatt ccccagcagg caggtaatta acagtgtgac tccctttttg
     5821 ctgcgagtgg ggctgataca gagagatgtg gcactatgga gcccacgggg tcctggcact
     5881 gggtgcccac ggaggtcccc atgtgctgca gtgtcaccgc ctccgaggag acagtattgt
     5941 ccctgcggtg tccctgcagc tcagctctgt ccacagggcc acctccagtt tggaggggac
     6001 acaatgcagc cccgatgcaa cccatcctcg cagcatccca gggacaaaga ccccactgca
     6061 agaccgcaca cagggctggg tcccgctccc ctaatatcta cagtgcttct tgcatggccc
     6121 cttaatcaat gcagttaatc agcatgcgct catgcaccgc tctggagctg caaagcccct
     6181 cgcagcgctg ctcaccaaca ccgcgcaccg ccccggccca gcctgcagca cgcgctgcaa
     6241 acaggaaaga aacaaaatat tgcccaaatg taggcaaagg cattcggctg ccttgacctc
     6301 cgccgggccg ggccctgcct gactcagctc cttactcagc gctcgcttcc tccctccggc
     6361 tgccaccgcc gcagcgcaca ccctgacaaa gagtggccct taacgggctc tgaggtgcac
     6421 ccagcagtgc actcagcagt ccaagggccg gcctggaggt ttgcaccgct acgtgctgac
     6481 attagcattg aacttggccc tgggtagtgc tgcaggccgg gcggggtggg tgtagagagt
     6541 gcagcgcgcg ttgcacccgg tgccccttcc cctcccttgc atcccagcag gctgcacccc
     6601 agcaccaggc ccgtgcatgc atgctcctgg tgttattgca gcctggtgca tgcatgcgtc
     6661 ttgatggtgc agcgctgtgc atgcatcctc cttggtgtgt agcagtctag tgcatgcata
     6721 cccctcggtg ttattgctgc tctgtgcacg cacgctcatt gtatcacttc atcccagtgc
     6781 atgcactcac actggagcga ttgctgctcg gtgcacgcac actcattgta tcactgcagc
     6841 tcagtggctg cacgcacacc ggtgttattg ctgctcggtg cgtgcatgca catcagtgtc
     6901 gctgcagctc agtgcatgca cgctcattgc ccatcgctat ccctgcctct cctgctggcg
     6961 ctccccggga ggtgacttca aggggaccgc aggaccacct cgggggtggg gggagggctg
     7021 cacacgcgga ccccgctccc cctccccaac aaagcactgt ggaatcaaaa aggggggagg
     7081 ggggatggag gggcgcgtca cacccccgcc ccacaccctc acc
//
