LOCUS       sequence-263613-        6140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Xenopus Kinesin-5 KSP 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 6140)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6140)
  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            "Unpublished"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: KSP_K560GFP
FEATURES             Location/Qualifiers
     source          1..6140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1588..1596
                     /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      1796..1805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1802..1804,1805..1807,1808..2518)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1802..1804,1805..1807,1808..2518)
                     /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(1847..1868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2108..2127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2455..2476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2528..2545
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2568..2585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2638..2656)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2652..2699
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2736..3191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2823..2842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3033..3054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3217..3321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3322..3390,3391..4182)
                     /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(3540..3559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4353..4941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4842..4861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5095..5112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5127..5266
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5192..5211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5352..5374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5368..5559)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6068..6087
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6068..6086
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6118..6140
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatacatatg gccagcaaga aggaggacaa agggaaaaac atccaggttg tggtccgatg
       61 ccggcccttc aatcagttgg aacgcaaggc gagctctcac tctgtgttgg aatgtgattc
      121 tcagaggaaa gaagtctatg ttcggaccgg aggaataaac gacaaactgg gaaagaaaac
      181 ctatacattt gacatggtgt ttgggcctgc agccaagcaa attgatgtgt acagaagcgt
      241 tgtgtgcccc atcttggatg aagttattat gggatataac tgcacaatct ttgcgtatgg
      301 gcagactggc acaggcaaga cctttactat ggaaggggaa agatcatcag atgaagagtt
      361 tacctgggaa caggaccctc tggcaggaat tattcctcga acgctgcatc agatatttga
      421 gaagctctct gagaacggca cagagttttc ggtgaaagtc tctctcctgg aaatctacaa
      481 cgaagaactc tttgatctgt taagcccatc tcccgatgtt ggggaaaggc tgcaaatgtt
      541 cgatgatcct cggaacaaaa gaggagtcat tatcaaaggg ctggaggaga tatctgtgca
      601 taacaaggat gaggtgtacc atatattgga gagaggtgca gccaggagga aaactgcttc
      661 tactctgatg aatgcatact ccagtcgatc ccattccgtg ttcgcggtca ccattcacat
      721 gaaagaaact actgttgatg gagaagagct tgtgaagatc gggaagctga acttggtgga
      781 tctggcggga agtgaaaata ttggtcgttc tggagctgtg gacaaaagag cacgcgaagc
      841 tggaaatatc aaccaatccc tgctcacctt gggcagggta ataactgctc tggtggagag
      901 aactcctcat attccataca gagaatccaa actcacaagg atcctgcagg attctcttgg
      961 agggaggaca aaaacctcta tcattgcaac agtgtcacct gcatccatta atctggagga
     1021 aactgtgagc actttggact atgccaacag agcaaagagt attatgaata agccagaggt
     1081 taaccagaaa cttacaaaga aggcgctagc cgaggaatgg aagcgacgct atgaaaagga
     1141 gaaggaaaag aacgcccgac taaagggtaa ggtggagaag cttgagatcg agcttgcgcg
     1201 ctggagagcg ggtgaaactg ttaaggcgga ggagcaaatc aacatggagg atctcatgga
     1261 ggcaagcacg cccaacctgg aagtggaggc agcacagacg gcggcggccg aggccgcttt
     1321 ggccgcccag cgaacggctc tcgccaatat gtccgcatcg gttgccgtga acgagcaggc
     1381 caggctggct acagagtgcg agcgtctcta ccagcagctg gacgacaagg atgaggagat
     1441 caatcagcag agccagtacg ccgagcagct caaggagcag gtgatggagc aggaggaact
     1501 catcgctaac gctcggcgtg agtatgagac tttgcagtcg gagatggcgc gaattcaaca
     1561 ggagaacgag tccgccaagg aagaggttaa ggaggtgctc caagctctcg aagagctgac
     1621 tgtaaactac gaccagaaat cccaggagat cgataacaag aacaaggata tcgatgccct
     1681 caacgaggag ctgcagcaga agcagtctgt gttcaacgcc gcctccacag agctacagca
     1741 gctcaagatg cttgttccgc gtgggcccgt gcaccgccta ggggatccac cggtcgccac
     1801 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1861 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1921 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1981 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2041 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2101 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2161 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2221 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2281 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2341 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2401 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2461 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagct
     2521 cgagagccat catcatcatc atcactaagc ggccgcactc gatcgagcac caccaccacc
     2581 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2641 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2701 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     2761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3181 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3421 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3481 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3541 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3601 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3661 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3721 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3781 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3841 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     3901 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3961 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4021 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4081 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4141 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5041 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5101 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5161 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5221 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     5281 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5341 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5401 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5461 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5521 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     5581 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     5641 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     5701 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     5761 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     5821 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5881 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5941 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     6001 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tctcgatccc
     6061 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
     6121 gtttaacttt aagaaggaga
//
