LOCUS       sequence-263587-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Drosophila Kinesin-1 full-length for microtubule gliding 
            assays.
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  JOURNAL   Unpublished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            "Unpublished"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: K980
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1542..1550
                     /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             2956..2973
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2974..2991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3014..3031
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3084..3102)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3098..3145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3182..3637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3269..3288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3479..3500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3663..3767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3768..3836,3837..4628)
                     /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(3986..4005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4799..5387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5288..5307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5541..5558
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5573..5712
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5638..5657)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5798..5820
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5814..6005)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6514..6533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6514..6532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atgagcgcag aacgagaaat tccagccgag
       61 gacagcatca aagtggtctg ccgattccga ccgctgaacg acagcgaaga gaaggccggc
      121 tccaagttcg tggtcaagtt ccccaacaat gtggaggaga actgcatatc catagcgggc
      181 aaggtgtatt tgttcgacaa ggtcttcaaa ccgaatgcat cccaggaaaa ggtctacaat
      241 gaggcggcca agtccattgt tacggatgtc ctggccgggt acaatggaac gatattcgca
      301 tacggtcaga cgtcctccgg aaaaacgcat acgatggagg gcgtgatcgg ggactccgta
      361 aaacagggta tcatacctag gatcgtcaac gacattttca atcacatcta cgcgatggag
      421 gtgaacctgg agtttcacat caaggtctcc tactacgaga tctacatgga caagattcga
      481 gatctgttgg acgtctccaa ggtgaacctt agtgtgcacg aggataagaa ccgggtgccg
      541 tacgtcaagg gcgctacgga acggttcgtc tcgtcgccgg aggatgtttt cgaggtgatc
      601 gaggagggca aatccaatcg tcacatcgct gtgacaaaca tgaacgagca ttcttcgcga
      661 tcccactcag tattccttat caatgtgaag caggagaacc tggagaacca gaagaaacta
      721 tccggcaaac tctacctggt ggatttggcc ggatccgaga aggtttccaa gactggagcg
      781 gagggaaccg ttcttgatga agccaagaac atcaacaagt cgctgtcggc cttgggcaac
      841 gtaatttctg ccctggcgga cggaaacaaa acgcacatcc cctaccgtga ttccaagcta
      901 acgcgcatcc tgcaggagtc gctgggaggc aacgcacgca caaccatcgt catctgctgc
      961 tctccagcca gtttcaacga atctgaaacg aagtcaacgc tggacttcgg tcgtagagcc
     1021 aagacagtga agaacgtggt ctgcgttaac gaggagctta ctgccgagga atggaagcga
     1081 cgctatgaaa aggagaagga aaagaacgcc cgactaaagg gtaaggtgga gaagcttgag
     1141 atcgagcttg cgcgctggag agcgggtgaa actgttaagg cggaggagca aatcaacatg
     1201 gaggatctca tggaggcaag cacgcccaac ctggaagtgg aggcagcaca gacggcggcg
     1261 gccgaggccg ctttggccgc ccagcgaacg gctctcgcca atatgtccgc atcggttgcc
     1321 gtgaacgagc aggccaggct ggctacagag tgcgagcgtc tctaccagca gctggacgac
     1381 aaggatgagg agatcaatca gcagagccag tacgccgagc agctcaagga gcaggtgatg
     1441 gagcaggagg aactcatcgc taacgctcgg cgtgagtatg agactttgca gtcggagatg
     1501 gcgcgaattc aacaggagaa cgagtccgcc aaggaagagg ttaaggaggt gctccaagct
     1561 ctcgaagagc tgactgtaaa ctacgaccag aaatcccagg agatcgataa caagaacaag
     1621 gatatcgatg ccctcaacga ggagctgcag cagaagcagt ctgtgttcaa cgccgcctcc
     1681 acagagctac agcagctcaa ggacatgtcc tcacaccaga agaagcgcat cacggaaatg
     1741 ctaaccaacc tactgcgcga cctcggcgaa gtgggccagg ccattgcccc cggcgagtcc
     1801 agcatcgacc ttaagatgag tgctctggct ggcacggatg ccagcaaggt ggaggaagat
     1861 ttcaccatgg cgcgtttgtt tatcagcaag atgaagacgg aggccaagaa cattgcccag
     1921 cgatgctcca acatggaaac acagcaggct gactccaaca agaagatctc cgaatatgag
     1981 aaagatctgg gcgagtaccg gctactcatt tcgcagcacg aggcacgcat gaagtcgctg
     2041 caggagtcga tgcgggaggc agagaacaag aagcgcacgc tcgaggaaca aatcgattcg
     2101 ctgcgcgagg aatgcgccaa gctcaaggcc gccgagcacg tttccgccgt taacgccgag
     2161 gagaaacagc gggctgagga gctgcgctcc atgttcgatt ctcagatgga cgagctacgc
     2221 gaagcccaca cccggcaggt gtccgagctc cgggacgaaa ttgccgccaa gcagcacgaa
     2281 atggacgaga tgaaggatgt ccatcaaaag ctgctcttgg cgcaccaaca gatgacggcc
     2341 gactacgaga aggtgcgcca ggaggatgcc gagaagtcca gcgagcttca gaacatcatc
     2401 ctcaccaacg agcgtcggga gcaagcgcgc aaagacctca agggcctgga ggacacggtg
     2461 gccaaggagt tgcagacgct acacaacctg cgaaaacttt tcgttcagga tctacagcaa
     2521 cgaatccgaa agaatgtcgt aaacgaggag agcgaggagg acggtggatc actcgcgcag
     2581 aaacagaaga tttccttctt ggagaacaac cttgatcagc tgaccaaggt gcacaagcaa
     2641 ttggtgcggg acaacgccga tctgcggtgc gagctgccca agctggagaa gcgtctacgc
     2701 tgtaccatgg agcgggtgaa agctctggag acagcgctca aggaggcgaa ggagggcgca
     2761 atgcgggatc gcaagcgcta ccaatacgag gtggaccgca tcaaggaggc ggtgcgacag
     2821 aagcatctgg gcagacgtgg cccacaggca cagatcgcaa agccgatccg gtccggccaa
     2881 ggtgcaatcg ccattcgtgg tggtggcgcc gttggaggac catccccgct ggcccaggtt
     2941 aatcctgtca actcgcttgt tccgcgtggg agccatcatc atcatcatca ctaagcggcc
     3001 gcactcgatc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     3061 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     3121 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     3181 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3361 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3481 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3601 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     3661 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3721 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3781 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3841 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3901 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3961 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4021 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4081 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4141 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4201 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4261 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     4321 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4381 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4441 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4501 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4561 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4621 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4681 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4921 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5461 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5521 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5581 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     5641 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     5701 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5761 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5821 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     5881 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     5941 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     6001 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     6061 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     6121 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     6181 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     6241 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     6301 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     6361 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     6421 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     6481 ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata gggagaccac
     6541 aacggtttcc ctctagaaat aat
//
