LOCUS       sequence-263530-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Kinesin-2 KIF3A head and neck linker, dimerized 
            through kinesin-1 neck-coil and coil-1 to 560 and GFP tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6117)
  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: 
            "Unpublished"
COMMENT     Sequence Label: KIF3A560GFP
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1565..1573
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      1773..1782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1779..1781,1782..1784,1785..2495)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1779..1781,1782..1784,1785..2495)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1824..1845)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2085..2104)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2432..2453
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2505..2522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2545..2562
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2615..2633)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2629..2676
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2713..3168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2800..2819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3010..3031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3194..3298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3299..3367,3368..4159)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3517..3536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4330..4918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4819..4838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5072..5089
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5104..5243
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5169..5188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5329..5351
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5345..5536)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6045..6064
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6045..6063
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6095..6117
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatacataat gccgatcaat aagtcggaga agccggaaag ctgcgataat gtgaaggtgg
       61 tggttaggtg ccggcctctc aatgagagag agaagtcaat gtgctataga caggccgtca
      121 gcgtggatga aatgagggga actatcaccg tccataagac cgattcctcc aacgaacctc
      181 caaagacatt tactttcgat actgtttttg gaccagaaag taaacaactt gatgtgtata
      241 acttaactgc aagaccaatt attgattctg ttctggaagg ctacaatggg accatctttg
      301 catatggaca gaccggaaca ggcaagactt tcaccatgga aggcgttcga gcagtaccgg
      361 ggctcagagg ggtcatcccc aactcatttg ctcacatatt tggtcacatc gcaaaagcag
      421 agggcgacac aaggtttttg gttcgagtgt catacttgga gatatataat gaagaagttc
      481 gagacctttt gggtaaagat cagacccaga ggttagaggt taaagaaagg cctgatgtgg
      541 gagtatatat caaagactta tcagcatatg tagtcaacaa cgctgacgac atggatagaa
      601 tcatgactct aggccacaaa aaccgttctg tcggtgcgac taatatgaac gagcacagct
      661 cccgatccca cgccatcttt acaattacta tagagtgcag tgagaaaggc gtggacggga
      721 acatgcacgt caggatgggc aagctccacc ttgtggatct ggctgggtca gaaagacaag
      781 caaaaactgg agccacagga cagcgcctca aggaagctac caaaatcaac ctttcattgt
      841 ctaccctggg caacgttatt tctgccttgg tggatggaaa aagcactcac gtgccgtatc
      901 ggaactccaa actgacccgt ctcctgcagg actccttagg agggaactca aaaaccatga
      961 tgtgtgcaaa tattgggcca gcagattata attatgatga aactatcagc accttgcggt
     1021 atgccaaccg cgccaagaac attaaaaaca aagcgagaat caatgaggac ccaaaggatg
     1081 cgctagccga ggaatggaag cgacgctatg aaaaggagaa ggaaaagaac gcccgactaa
     1141 agggtaaggt ggagaagctt gagatcgagc ttgcgcgctg gagagcgggt gaaactgtta
     1201 aggcggagga gcaaatcaac atggaggatc tcatggaggc aagcacgccc aacctggaag
     1261 tggaggcagc acagacggcg gcggccgagg ccgctttggc cgcccagcga acggctctcg
     1321 ccaatatgtc cgcatcggtt gccgtgaacg agcaggccag gctggctaca gagtgcgagc
     1381 gtctctacca gcagctggac gacaaggatg aggagatcaa tcagcagagc cagtacgccg
     1441 agcagctcaa ggagcaggtg atggagcagg aggaactcat cgctaacgct cggcgtgagt
     1501 atgagacttt gcagtcggag atggcgcgaa ttcaacagga gaacgagtcc gccaaggaag
     1561 aggttaagga ggtgctccaa gctctcgaag agctgactgt aaactacgac cagaaatccc
     1621 aggagatcga taacaagaac aaggatatcg atgccctcaa cgaggagctg cagcagaagc
     1681 agtctgtgtt caacgccgcc tccacagagc tacagcagct caagatgctt gttccgcgtg
     1741 ggcccgtgca ccgcctaggg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1801 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1861 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1921 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1981 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2041 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2101 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2161 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2221 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2281 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2341 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2401 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2461 ccgggatcac tctcggcatg gacgagctgt acaagctcga gagccatcat catcatcatc
     2521 actaagcggc cgcactcgat cgagcaccac caccaccacc actgagatcc ggctgctaac
     2581 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2641 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2701 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2761 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2821 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2881 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2941 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3001 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3061 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3121 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3301 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3361 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3421 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3481 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3541 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3601 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3661 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3721 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3781 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3841 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3901 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3961 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4021 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4081 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4141 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4441 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4981 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5041 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5101 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     5161 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     5221 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     5281 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5341 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     5401 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     5461 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     5521 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     5581 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     5641 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     5701 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     5761 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     5821 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     5881 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     5941 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     6001 gtggccagga cccaacgctg cccgagatct cgatcccgcg aaattaatac gactcactat
     6061 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggaga
//
