LOCUS       sequence-380528-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP-tagged Kif 3B CC 391-592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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 CC 391-592
FEATURES             Location/Qualifiers
     source          1..7374
                     /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    2793..2927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2830..2849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2884..2903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2958..2974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2958..2974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2971..2993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2982..2998
                     /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(3006..3012,3013..3030,3031..3036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3051..3072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3189..3206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3360..3948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3440..3459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4119..4910,4911..4979))
                     /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     4742..4761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4980..5084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5152..5170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5208..5230)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5330..5349
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5543..5565
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5557..5574
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5558..5574
                     /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 caaggggggt ggggaagagg aggaggagga gggagaagag
     2101 ggtgaggagg acggggatga taaggatgat tactggcggg aacagcaaga aaaactggag
     2161 attgagaagc gggccattgt agaggaccac agcttggttg cagaggagaa gatgaggctg
     2221 ctgaaggaga aggagaaaaa gatggaggac ctgcggcgag agaaggatgc tgcagagatg
     2281 ctgggtgcca aaatcaaggc catggagagt aagctgcttg tcggaggaaa aaatatagta
     2341 gatcatacaa acgaacagca gaagatcctg gagcaaaaac gccaggagat cgcagagcag
     2401 aaacgtcgag aaagagaaat ccagcagcaa atggagagtc gagatgagga gaccctggaa
     2461 ctgaaagaga cttacacctc actgcagcag gaggtagaca tcaagaccaa aaagctcaaa
     2521 aagctctttt ccaagcttca ggccgtgaag gccgagatcc acgacctcca agaagaacac
     2581 atcaaagagc gccaggagct ggagcagacg cagaatgaac tcacccggga gctaaagctc
     2641 aagcatctta ttatagaaaa ctttatttaa aatctagata agtaatgatc ataatcagcc
     2701 atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2761 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc gggtaccgag
     2941 ctcgaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3001 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3061 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3121 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3181 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3241 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3301 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3361 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3421 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3481 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3541 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3601 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3661 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3721 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3781 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3841 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3901 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3961 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4021 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4081 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4141 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4201 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4261 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4321 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4381 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4441 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4501 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4561 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4621 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4681 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4741 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4801 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4861 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4921 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4981 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5041 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5101 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5161 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5221 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5281 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     5341 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5401 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     5461 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     5521 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     5581 ttgggatctt tgcattggcc cacggctctc aggatgggga tgctcccctt cagcacccgg
     5641 ttccccttgg aaactgatgg tcctggctct gtggcatggc agtggcactg tgaggagccc
     5701 cctaccagca gcacacagtg gggttggcac tgccacgctc cggatgccgc gctctgatcc
     5761 aaccccataa tcaagggaac ccgaattgcc ccatcattgc ccccaccacc cccatcctgc
     5821 cgggccctca caccccacgc tgccttctgg tgacattccc cagcccaaac ccacggcttc
     5881 atggctaccg cggggcattt cccattgccg ccccattatc agctctgcac acctcccgct
     5941 gtacccatgc ctcgtggctg cccttctttg acgtataatc ttctaattaa tacccggcct
     6001 tgtcaaagtg gagcacaaac gttaattaat tccccagcag gcaggtaatt aacagtgtga
     6061 ctcccttttt gctgcgagtg gggctgatac agagagatgt ggcactatgg agcccacggg
     6121 gtcctggcac tgggtgccca cggaggtccc catgtgctgc agtgtcaccg cctccgagga
     6181 gacagtattg tccctgcggt gtccctgcag ctcagctctg tccacagggc cacctccagt
     6241 ttggagggga cacaatgcag ccccgatgca acccatcctc gcagcatccc agggacaaag
     6301 accccactgc aagaccgcac acagggctgg gtcccgctcc cctaatatct acagtgcttc
     6361 ttgcatggcc ccttaatcaa tgcagttaat cagcatgcgc tcatgcaccg ctctggagct
     6421 gcaaagcccc tcgcagcgct gctcaccaac accgcgcacc gccccggccc agcctgcagc
     6481 acgcgctgca aacaggaaag aaacaaaata ttgcccaaat gtaggcaaag gcattcggct
     6541 gccttgacct ccgccgggcc gggccctgcc tgactcagct ccttactcag cgctcgcttc
     6601 ctccctccgg ctgccaccgc cgcagcgcac accctgacaa agagtggccc ttaacgggct
     6661 ctgaggtgca cccagcagtg cactcagcag tccaagggcc ggcctggagg tttgcaccgc
     6721 tacgtgctga cattagcatt gaacttggcc ctgggtagtg ctgcaggccg ggcggggtgg
     6781 gtgtagagag tgcagcgcgc gttgcacccg gtgccccttc ccctcccttg catcccagca
     6841 ggctgcaccc cagcaccagg cccgtgcatg catgctcctg gtgttattgc agcctggtgc
     6901 atgcatgcgt cttgatggtg cagcgctgtg catgcatcct ccttggtgtg tagcagtcta
     6961 gtgcatgcat acccctcggt gttattgctg ctctgtgcac gcacgctcat tgtatcactt
     7021 catcccagtg catgcactca cactggagcg attgctgctc ggtgcacgca cactcattgt
     7081 atcactgcag ctcagtggct gcacgcacac cggtgttatt gctgctcggt gcgtgcatgc
     7141 acatcagtgt cgctgcagct cagtgcatgc acgctcattg cccatcgcta tccctgcctc
     7201 tcctgctggc gctccccggg aggtgacttc aaggggaccg caggaccacc tcgggggtgg
     7261 ggggagggct gcacacgcgg accccgctcc ccctccccaa caaagcactg tggaatcaaa
     7321 aaggggggag gggggatgga ggggcgcgtc acacccccgc cccacaccct cacc
//
