LOCUS       sequence-263572-        4907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Xenopus Kinesin-5 KSP head and neck linker, dimerized 
            through kinesin-1 neck-coil to 406 for biochemical assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4907)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4907)
  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            "Unpublished"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_K406
FEATURES             Location/Qualifiers
     source          1..4907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1368..1385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1408..1425
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1478..1496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1492..1539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1576..2031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1663..1682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1873..1894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2057..2161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2162..2230,2231..3022)
                     /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(2380..2399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3193..3781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3682..3701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3935..3952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3967..4106
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4032..4051)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4192..4214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4208..4399)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atggccagca agaaggagga caaagggaaa aacatccagg
      121 ttgtggtccg atgccggccc ttcaatcagt tggaacgcaa ggcgagctct cactctgtgt
      181 tggaatgtga ttctcagagg aaagaagtct atgttcggac cggaggaata aacgacaaac
      241 tgggaaagaa aacctataca tttgacatgg tgtttgggcc tgcagccaag caaattgatg
      301 tgtacagaag cgttgtgtgc cccatcttgg atgaagttat tatgggatat aactgcacaa
      361 tctttgcgta tgggcagact ggcacaggca agacctttac tatggaaggg gaaagatcat
      421 cagatgaaga gtttacctgg gaacaggacc ctctggcagg aattattcct cgaacgctgc
      481 atcagatatt tgagaagctc tctgagaacg gcacagagtt ttcggtgaaa gtctctctcc
      541 tggaaatcta caacgaagaa ctctttgatc tgttaagccc atctcccgat gttggggaaa
      601 ggctgcaaat gttcgatgat cctcggaaca aaagaggagt cattatcaaa gggctggagg
      661 agatatctgt gcataacaag gatgaggtgt accatatatt ggagagaggt gcagccagga
      721 ggaaaactgc ttctactctg atgaatgcat actccagtcg atcccattcc gtgttcgcgg
      781 tcaccattca catgaaagaa actactgttg atggagaaga gcttgtgaag atcgggaagc
      841 tgaacttggt ggatctggcg ggaagtgaaa atattggtcg ttctggagct gtggacaaaa
      901 gagcacgcga agctggaaat atcaaccaat ccctgctcac cttgggcagg gtaataactg
      961 ctctggtgga gagaactcct catattccat acagagaatc caaactcaca aggatcctgc
     1021 aggattctct tggagggagg acaaaaacct ctatcattgc aacagtgtca cctgcatcca
     1081 ttaatctgga ggaaactgtg agcactttgg actatgccaa cagagcaaag agtattatga
     1141 ataagccaga ggttaaccag aaacttacaa agaaggcgct agccgaggaa tggaagcgac
     1201 gctatgaaaa ggagaaggaa aagaacgccc gactaaaggg taaggtggag aagcttgaga
     1261 tcgagcttgc gcgctggaga gcgggtgaaa ctgttaaggc ggaggagcaa atcaacatgg
     1321 aggatctcat ggaggcaagc acgcccaacc tggaagtgga ggctagccat catcatcatc
     1381 atcactaagc ggccgcactc gatcgagcac caccaccacc accactgaga tccggctgct
     1441 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1501 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1561 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1621 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1681 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1741 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1801 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1861 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1921 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1981 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2041 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2101 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2161 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2221 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2281 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2341 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2401 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2461 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2521 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2581 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2641 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2701 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2761 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2821 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2881 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2941 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3001 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3061 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3301 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3841 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3901 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3961 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     4021 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4081 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4141 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4201 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4261 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4321 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4381 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4441 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4501 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4561 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4621 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4681 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4741 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4801 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     4861 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaat
//
