LOCUS       sequence-263579-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Drosophila Kinesin-1 with rigor mutation 
            (R210A) for adhering microtubules to surfaces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6563)
  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            "Unpublished"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: KRigor980
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1512..1520
                     /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)"
     CDS             2926..2943
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2944..2961
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2984..3001
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3054..3072)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3068..3115
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3152..3607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3239..3258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3449..3470
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3633..3737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3738..3806,3807..4598)
                     /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(3956..3975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4769..5357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5258..5277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5511..5528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5543..5682
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5608..5627)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5768..5790
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5784..5975)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6484..6503
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6484..6502
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6534..6556
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgagcgcag aacgagaaat tccagccgag gacagcatca aagtggtctg ccgattccga
       61 ccgctgaacg acagcgaaga gaaggccggc tccaagttcg tggtcaagtt ccccaacaat
      121 gtggaggaga actgcatatc catagcgggc aaggtgtatt tgttcgacaa ggtcttcaaa
      181 ccgaatgcat cccaggaaaa ggtctacaat gaggcggcca agtccattgt tacggatgtc
      241 ctggccgggt acaatggaac gatattcgca tacggtcaga cgtcctccgg aaaaacgcat
      301 acgatggagg gcgtgatcgg ggactccgta aaacagggta tcatacctag gatcgtcaac
      361 gacattttca atcacatcta cgcgatggag gtgaacctgg agtttcacat caaggtctcc
      421 tactacgaga tctacatgga caagattcga gatctgttgg acgtctccaa ggtgaacctt
      481 agtgtgcacg aggataagaa ccgggtgccg tacgtcaagg gcgctacgga acggttcgtc
      541 tcgtcgccgg aggatgtttt cgaggtgatc gaggagggca aatccaatcg tcacatcgct
      601 gtgacaaaca tgaacgagca ttcttcggca tcccactcag tattccttat caatgtgaag
      661 caggagaacc tggagaacca gaagaaacta tccggcaaac tctacctggt ggatttggcc
      721 ggatccgaga aggtttccaa gactggagcg gagggaaccg ttcttgatga agccaagaac
      781 atcaacaagt cgctgtcggc cttgggcaac gtaatttctg ccctggcgga cggaaacaaa
      841 acgcacatcc cctaccgtga ttccaagcta acgcgcatcc tgcaggagtc gctgggaggc
      901 aacgcacgca caaccatcgt catctgctgc tctccagcca gtttcaacga atctgaaacg
      961 aagtcaacgc tggacttcgg tcgtagagcc aagacagtga agaacgtggt ctgcgttaac
     1021 gaggagctta ctgccgagga atggaagcga cgctatgaaa aggagaagga aaagaacgcc
     1081 cgactaaagg gtaaggtgga gaagcttgag atcgagcttg cgcgctggag agcgggtgaa
     1141 actgttaagg cggaggagca aatcaacatg gaggatctca tggaggcaag cacgcccaac
     1201 ctggaagtgg aggcagcaca gacggcggcg gccgaggccg ctttggccgc ccagcgaacg
     1261 gctctcgcca atatgtccgc atcggttgcc gtgaacgagc aggccaggct ggctacagag
     1321 tgcgagcgtc tctaccagca gctggacgac aaggatgagg agatcaatca gcagagccag
     1381 tacgccgagc agctcaagga gcaggtgatg gagcaggagg aactcatcgc taacgctcgg
     1441 cgtgagtatg agactttgca gtcggagatg gcgcgaattc aacaggagaa cgagtccgcc
     1501 aaggaagagg ttaaggaggt gctccaagct ctcgaagagc tgactgtaaa ctacgaccag
     1561 aaatcccagg agatcgataa caagaacaag gatatcgatg ccctcaacga ggagctgcag
     1621 cagaagcagt ctgtgttcaa cgccgcctcc acagagctac agcagctcaa ggacatgtcc
     1681 tcacaccaga agaagcgcat cacggaaatg ctaaccaacc tactgcgcga cctcggcgaa
     1741 gtgggccagg ccattgcccc cggcgagtcc agcatcgacc ttaagatgag tgctctggct
     1801 ggcacggatg ccagcaaggt ggaggaagat ttcaccatgg cgcgtttgtt tatcagcaag
     1861 atgaagacgg aggccaagaa cattgcccag cgatgctcca acatggaaac acagcaggct
     1921 gactccaaca agaagatctc cgaatatgag aaagatctgg gcgagtaccg gctactcatt
     1981 tcgcagcacg aggcacgcat gaagtcgctg caggagtcga tgcgggaggc agagaacaag
     2041 aagcgcacgc tcgaggaaca aatcgattcg ctgcgcgagg aatgcgccaa gctcaaggcc
     2101 gccgagcacg tttccgccgt taacgccgag gagaaacagc gggctgagga gctgcgctcc
     2161 atgttcgatt ctcagatgga cgagctacgc gaagcccaca cccggcaggt gtccgagctc
     2221 cgggacgaaa ttgccgccaa gcagcacgaa atggacgaga tgaaggatgt ccatcaaaag
     2281 ctgctcttgg cgcaccaaca gatgacggcc gactacgaga aggtgcgcca ggaggatgcc
     2341 gagaagtcca gcgagcttca gaacatcatc ctcaccaacg agcgtcggga gcaagcgcgc
     2401 aaagacctca agggcctgga ggacacggtg gccaaggagt tgcagacgct acacaacctg
     2461 cgaaaacttt tcgttcagga tctacagcaa cgaatccgaa agaatgtcgt aaacgaggag
     2521 agcgaggagg acggtggatc actcgcgcag aaacagaaga tttccttctt ggagaacaac
     2581 cttgatcagc tgaccaaggt gcacaagcaa ttggtgcggg acaacgccga tctgcggtgc
     2641 gagctgccca agctggagaa gcgtctacgc tgtaccatgg agcgggtgaa agctctggag
     2701 acagcgctca aggaggcgaa ggagggcgca atgcgggatc gcaagcgcta ccaatacgag
     2761 gtggaccgca tcaaggaggc ggtgcgacag aagcatctgg gcagacgtgg cccacaggca
     2821 cagatcgcaa agccgatccg gtccggccaa ggtgcaatcg ccattcgtgg tggtggcgcc
     2881 gttggaggac catccccgct ggcccaggtt aatcctgtca actcgcttgt tccgcgtggg
     2941 agccatcatc atcatcatca ctaagcggcc gcactcgatc gagcaccacc accaccacca
     3001 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3061 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     3121 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3601 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3661 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4261 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5521 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5581 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5641 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     5701 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5761 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     5821 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     5881 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     5941 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     6001 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     6061 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     6121 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     6181 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     6241 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     6301 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     6361 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     6421 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga
     6481 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt
     6541 aactttaaga aggagatata cat
//
