LOCUS       sequence-380543-        7237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3B DD 593-747.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  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 7237)
  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-KIF3B DD 593-747
FEATURES             Location/Qualifiers
     source          1..7237
                     /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    2656..2790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2747..2766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2821..2837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2821..2837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2834..2856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2845..2861
                     /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(2869..2875,2876..2893,2894..2899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2914..2935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3052..3069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3303..3322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3982..4773,4774..4842))
                     /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     4605..4624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4843..4947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5015..5033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5071..5093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5193..5212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5406..5428
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5420..5437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5421..5437
                     /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 caaacccttg gaagagaaga ataaaattat gaatagatcc
     2101 ttttttgatg atgaagaaga ccattggaaa ttacatccta taaccagact ggaaaaccag
     2161 cagatgatga agcggcctgt ctctgctgtg ggatacaaga gacctctgag ccagcatgca
     2221 cggatgtcca tgatgattcg gccagagccc cggtacaggg cggagaacat catgctcttg
     2281 gagttagata tgcccagccg gacaaccaga gactacgagg gcccagccat ctctcccaag
     2341 gttcaggctg cgctggatgc agctctgcag gatgaagatg agatacaggt ggatgcctca
     2401 tcctttgaaa gtactgccag cagaaaaccc aaggccaggc ccaagagtgg caggaaatca
     2461 ggatcctcct cttcctcctc aggaaaccct gcatctcagt tttacccaca gtctcggggg
     2521 ctggttccca agtaatctag ataagtaatg atcataatca gccatatcac atctgtagag
     2581 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2641 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2761 ctcatcaatg tatcttatca tgtctggatc cccgggtacc gagctcgaat tcgtaatcat
     2821 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2881 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2941 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3001 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4981 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5041 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5101 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5161 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5221 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5281 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5341 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5401 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagttgggat ctttgcattg
     5461 gcccacggct ctcaggatgg ggatgctccc cttcagcacc cggttcccct tggaaactga
     5521 tggtcctggc tctgtggcat ggcagtggca ctgtgaggag ccccctacca gcagcacaca
     5581 gtggggttgg cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg
     5641 aacccgaatt gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca
     5701 cgctgccttc tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca
     5761 tttcccattg ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg
     5821 ctgcccttct ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca
     5881 aacgttaatt aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga
     5941 gtggggctga tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc
     6001 ccacggaggt ccccatgtgc tgcagtgtca ccgcctccga ggagacagta ttgtccctgc
     6061 ggtgtccctg cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg
     6121 cagccccgat gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg
     6181 cacacagggc tgggtcccgc tcccctaata tctacagtgc ttcttgcatg gccccttaat
     6241 caatgcagtt aatcagcatg cgctcatgca ccgctctgga gctgcaaagc ccctcgcagc
     6301 gctgctcacc aacaccgcgc accgccccgg cccagcctgc agcacgcgct gcaaacagga
     6361 aagaaacaaa atattgccca aatgtaggca aaggcattcg gctgccttga cctccgccgg
     6421 gccgggccct gcctgactca gctccttact cagcgctcgc ttcctccctc cggctgccac
     6481 cgccgcagcg cacaccctga caaagagtgg cccttaacgg gctctgaggt gcacccagca
     6541 gtgcactcag cagtccaagg gccggcctgg aggtttgcac cgctacgtgc tgacattagc
     6601 attgaacttg gccctgggta gtgctgcagg ccgggcgggg tgggtgtaga gagtgcagcg
     6661 cgcgttgcac ccggtgcccc ttcccctccc ttgcatccca gcaggctgca ccccagcacc
     6721 aggcccgtgc atgcatgctc ctggtgttat tgcagcctgg tgcatgcatg cgtcttgatg
     6781 gtgcagcgct gtgcatgcat cctccttggt gtgtagcagt ctagtgcatg catacccctc
     6841 ggtgttattg ctgctctgtg cacgcacgct cattgtatca cttcatccca gtgcatgcac
     6901 tcacactgga gcgattgctg ctcggtgcac gcacactcat tgtatcactg cagctcagtg
     6961 gctgcacgca caccggtgtt attgctgctc ggtgcgtgca tgcacatcag tgtcgctgca
     7021 gctcagtgca tgcacgctca ttgcccatcg ctatccctgc ctctcctgct ggcgctcccc
     7081 gggaggtgac ttcaagggga ccgcaggacc acctcggggg tggggggagg gctgcacacg
     7141 cggaccccgc tccccctccc caacaaagca ctgtggaatc aaaaaggggg gaggggggat
     7201 ggaggggcgc gtcacacccc cgccccacac cctcacc
//
