LOCUS       sequence-264857-        4721 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Xenopus Kinesin-5 KSP motor and neck linker domain monomer
            for biochemical assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4721)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4721)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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_HNL
FEATURES             Location/Qualifiers
     source          1..4721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     289..308
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        289..307
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             339..361
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1470..1487
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1510..1527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1580..1598)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1594..1641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1678..2133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1765..1784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1975..1996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2159..2263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2264..2332,2333..3124)
                     /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(2482..2501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3295..3883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3784..3803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4037..4054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4069..4208
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4134..4153)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4294..4316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4310..4501)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
       61 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
      121 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
      181 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
      241 acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta atacgactca
      301 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
      361 atatacatat ggccagcaag aaggaggaca aagggaaaaa catccaggtt gtggtccgat
      421 gccggccctt caatcagttg gaacgcaagg cgagctctca ctctgtgttg gaatgtgatt
      481 ctcagaggaa agaagtctat gttcggaccg gaggaataaa cgacaaactg ggaaagaaaa
      541 cctatacatt tgacatggtg tttgggcctg cagccaagca aattgatgtg tacagaagcg
      601 ttgtgtgccc catcttggat gaagttatta tgggatataa ctgcacaatc tttgcgtatg
      661 ggcagactgg cacaggcaag acctttacta tggaagggga aagatcatca gatgaagagt
      721 ttacctggga acaggaccct ctggcaggaa ttattcctcg aacgctgcat cagatatttg
      781 agaagctctc tgagaacggc acagagtttt cggtgaaagt ctctctcctg gaaatctaca
      841 acgaagaact ctttgatctg ttaagcccat ctcccgatgt tggggaaagg ctgcaaatgt
      901 tcgatgatcc tcggaacaaa agaggagtca ttatcaaagg gctggaggag atatctgtgc
      961 ataacaagga tgaggtgtac catatattgg agagaggtgc agccaggagg aaaactgctt
     1021 ctactctgat gaatgcatac tccagtcgat cccattccgt gttcgcggtc accattcaca
     1081 tgaaagaaac tactgttgat ggagaagagc ttgtgaagat cgggaagctg aacttggtgg
     1141 atctggcggg aagtgaaaat attggtcgtt ctggagctgt ggacaaaaga gcacgcgaag
     1201 ctggaaatat caaccaatcc ctgctcacct tgggcagggt aataactgct ctggtggaga
     1261 gaactcctca tattccatac agagaatcca aactcacaag gatcctgcag gattctcttg
     1321 gagggaggac aaaaacctct atcattgcaa cagtgtcacc tgcatccatt aatctggagg
     1381 aaactgtgag cactttggac tatgccaaca gagcaaagag tattatgaat aagccagagg
     1441 ttaaccagaa acttacaaag aaggcgctac atcatcatca tcatcactaa gcggccgcac
     1501 tcgatcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     1561 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1621 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     1681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1981 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2041 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2101 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3421 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3961 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4021 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4081 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     4141 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     4201 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4261 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4321 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4381 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4441 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4501 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     4561 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     4621 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     4681 gccagcgctt cgttaataca gatgtaggtg ttccacaggg t
//
