LOCUS       sequence-414438-        7888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses wild-type Octopus kinesin-1 motor with HaloTag in 
            Drosophila S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7888)
  AUTHORS   Birk MA, Liscovitch-Brauer N, Dominguez MJ, McNeme S, Yue Y, Hoff 
            JD, Twersky I, Verhey KJ, Sutton RB, Eisenberg E, Rosenthal JJC
  TITLE     Temperature-dependent RNA editing in octopus extensively recodes the
            neural proteome.
  JOURNAL   Cell. 2023 Jun 8;186(12):2544-2555.e13. doi: 
            10.1016/j.cell.2023.05.004. Epub 2023 Jun 8.
  PUBMED    37295402
REFERENCE   2  (bases 1 to 7888)
  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"; doi: 
            "10.1016/j.cell.2023.05.004"; journalName: "Cell"; date: "2023-06-8-
            8"; volume: "186"; issue: "12"; pages: "2544-2555.e13"
COMMENT     Sequence Label: pAc5.1-V5-HisB-ObKHC-wt-HaloTag
FEATURES             Location/Qualifiers
     source          1..7888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..2562
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             4293..5183
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             5191..5232
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5242..5259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5282..5299)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5332..5466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5356..5375)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5410..5429
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6147..6166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6826..7617,7618..7686))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7449..7468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7687..7791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7859..7877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caagaattaa ttctatattc taaaaacaca aatgatactt ctaaaaaaaa atcatgaatg
       61 gcatcaactc tgaatcaaat ctttgcagat gcacctactt ctcatttcca ctgtcacatc
      121 atttttccag atctcgctgc ctgttatgtg gcccacaaac caagacacgt tttatggcca
      181 ttaaagctgg ctgatcgtcg ccaaacacca aatacataat gaatatgtac acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg gagtaggaga atgcggagga gaaggagaac
      301 gagctgatct agtatctctc cacaatccaa tgccaactga ccaactggcc atattcggag
      361 caatttgaag ccaatttcca tcgcctggcg atcgctccat tcttggctat atgtttttca
      421 ccgttacccg gggccatttt caaagactcg tcggcaagat aagattgtgt cactcgctgt
      481 ctctcttcat ttgtcgaaga atgctgagga atttcgcgat gacgtcggcg agtattttga
      541 agaatgagaa taatttgtat ttatacgaaa atcagttagt ggaattttct acaaaaacat
      601 gttatctata gataattttg ttgcaaaata tgttgactat gacaaagatt gtatgtatat
      661 acctttaatg tattctcatt ttcttatgta tttataatgg caatgatgat actgatgata
      721 ttttaagatg atgccagacc aaaaggcttg aatttctgcg tcttttgccg aacgcagtgc
      781 atgtgcaatt gttgtttttt ggaatattca attttcggac tgtccgcttt gatttcagtt
      841 tcttggctta ttcaaaaagc aaagtaaagc caaaaaagcg agatggcaat accaaatgcg
      901 gcaaaacggt agtggaagga aaggggtgcg gggcagcgga aggaagggtg gggcggggcg
      961 tggcggggtc tgtggctggg cgcgacgtca ccgacgttgg agccactcct ttgaccatgt
     1021 gtgcgtgtgt gtattattcg tgtctcgcca ctcgccggtt gtttttttct ttttatgctg
     1081 cgctctctct agcgccatct cgcttacgca tgctcaacgc accgcatgtt gccgtttcct
     1141 tttatgcgtc attttggctc gaaataggca attatttaaa caaagattag tcaacgaaaa
     1201 cgctaaaata aataagtcta caatatggtt acttattgcc atgtgtgtgc agccaacgat
     1261 agcaacaaaa gcaacaacac aggtggcttt ccctctttca ctttttgttt gcaagccgcg
     1321 tgcgagcaag acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt
     1381 tggcgaaaaa catcaaggcg cctgatacga atgcatttgc aataacaatt gcgatattta
     1441 atattgttta tgaagctgtt tgacttcaaa acacacaaaa aaaaaaataa aacaaattat
     1501 ttgaaagaga attaggaatc ggacgcttat cgttagggta acaacaagaa atgcttactg
     1561 agtcacagcc tctggaaaac tgccgcaagc cagagagaga gagaaaaaga gggagagcag
     1621 cttagaccgc atgtgcttgt gtgtgaggcg tctctctctt cgtctctgtt gcgcaaacgc
     1681 atagactgca ctgaaaaaat cgattaccta ttttttatga atgaatattt gcactattac
     1741 tattcaaaac tattaagata gcaatcacat tcaatagcca aatactatac cacctgagcg
     1801 atgcaacgaa atgatcaatt tgagcaaaaa tgctgcatat ttaggacggc atcattatag
     1861 aaatgcttct tgctgtgtac ttttctctcg tctggcagct gtttcgccgt tattgttaaa
     1921 accggcttaa gttaggtgtg ttttctacga ctagtgaatg ccctactaga agatgtgtgt
     1981 tgcacaaaat gtccctggaa taaccaattt gaagtgcaga tagcagtaaa cgtaagctaa
     2041 tatgaatatt atttaactgt aatgttttaa tatcgctgga cattactaat aaacccacta
     2101 taaacacatg tacatatgta tgttttggca tacaatgagt agttggggaa aaaatgtgta
     2161 aaagcaccgt gaccatcaca gcataaagat aaccagctga agtatcgaat atgagtaacc
     2221 cccaaattga atcacatgcc gcaactgata ggacccatgg aagtacactc ttcatggcga
     2281 tatacaagac acacacaagc acgaacaccc agttgcggag gaaattctcc gtaaatgaaa
     2341 acccaatcgg cgaacaattc atacccatat atggtaaaag ttttgaacgc gacttgagag
     2401 cggagagcat tgcggctgat aaggttttag cgctaagcgg gctttataaa acgggctgcg
     2461 ggaccagttt tcatatcact accgtttgag ttcttgtgct gtgtggatac tcctcccgac
     2521 acaaagccgc tccatcagcc agcagtcgtc taatccagag accccggatc ggggtaccta
     2581 ctagtccagt gtggtggatc aacatggatg catcttcaga atgcaatatt aaagtcatat
     2641 gccgggtgag acctctcaat gatgcagaag agcgagcagg ctccaagttt atactcaaat
     2701 ttcctaccga tgattctata agtatagctg gtaaagtgtt cgtttttgat aaagtgctca
     2761 agccaaacgt aagtcaagag tacgtctaca atgttggcgc acagccgatt gtggctgatg
     2821 tccttagtgg atataacggg actattttcg cctacggtca aaccagttcc ggaaagacgc
     2881 acacaatgga aggagtcctt gataaaccgt ctatgcacgg catcattccg cgcatagttc
     2941 aggacatatt caattatata tacgggatgg atgagaacct agagttccac ataaagattt
     3001 cctactatga aatatacttg gataaaataa gagatctcct cgatgtaact aaaacgaact
     3061 tgtcagtgca cgaggacaaa aatcgcgtac cgtatgtcaa gggagccact gagagatttg
     3121 tcagtagccc ggaggaggtt atggaagtta ttgatgaggg taaaaataac cggcatgtcg
     3181 ctgtaaccaa catgaacgag cacagttcaa ggtctcactc cgtgtttttg atcaacgtaa
     3241 aacaggagaa cgtcgaaacg caaaagaaac tttccggaaa actgtacctt gtggaccttg
     3301 ccggatcaga aaaggtgtct aaaacgggcg cggaaggagc ggttctcgac gaggcaaaaa
     3361 acattaataa gagtcttagc gcactgggga atgttatcag cgcgctcgct gatgggaaca
     3421 agagccatgt cccctaccga gatagtaaac tgacgcggat cttgcaagaa tctcttggag
     3481 gcaatgcaag gacaaccatg gtcatatgct gtagtcctgc ttcctacaat gaaagcgaaa
     3541 ccaaatcaac actcctgttc ggtcaaaggg cgaaaactat caagaatgtt gtcagtgtaa
     3601 acgaagaatt gacggccgac gagtggaagc gcagatatga gaaggaaaag gagaaatgca
     3661 cgaaattgaa agctacgctc gccagacttg aatctgaact gcaaaaatgg cgaacaggcc
     3721 aacccgtatc tgttgatgac caggtggatc tcaaggctga tatacctgcg gaatcccccg
     3781 cgacaagtac cacatccttg gctgggaata tggtggtatc aatgaatgaa ggagaccgct
     3841 cacaactgga ggaggagcga ttgaagctgt atcagcaact cgacgacaag gatgacgaga
     3901 tcaataatca gagccagctc atagagaaat tgaatgaaca aatgatggag caagaagaac
     3961 tcatatccca atcacggaag gattacgaga atcttcagca agagatgagc cgaatccaga
     4021 gcgacaatga gtctgctaag gacgaagtga aagaagtgct gcaagcattg gaggaattgg
     4081 cgatgaacta tgaccaaaag tcccaggaag tggaaaacaa gaataaggag aatgagaatc
     4141 tctccgatga gttgtcccaa aaacttagta cccttaattc actccagaat gaactggatc
     4201 aactaaagga cagcagtatg catcaccgca agcgcgtgac cgacatgatg atcaaccttc
     4261 tcaaggacct gggagatgga ggtggaggtt caatggcgga aataggcacg ggctttccgt
     4321 ttgaccctca ctacgttgag gtacttgggg aacggatgca ttacgtagat gtagggccaa
     4381 gagacgggac gccggtactc ttcctgcacg ggaatccaac gagcagctac gtgtggagaa
     4441 atatcatccc gcatgtcgcc ccgactcata ggtgtatagc tccagatttg attggcatgg
     4501 gaaaatcaga taaacctgat cttggttact tttttgacga ccatgtgagg tttatggacg
     4561 cattcataga ggcgcttggt ctcgaggagg ttgtacttgt catacatgac tggggttctg
     4621 cactcggctt ccactgggca aagcgcaatc ctgaaagggt aaaaggcatc gcatttatgg
     4681 agtttattcg gcctattccc acgtgggatg aatggccgga atttgctaga gagacgtttc
     4741 aggcatttcg caccacggat gtagggcgga aacttattat tgaccaaaat gtattcatag
     4801 aggggactct tccgatgggt gttgtgaggc cgcttactga ggtggaaatg gatcactata
     4861 gagagccttt ccttaatcca gtcgaccgag agcctctgtg gcggttcccc aatgaacttc
     4921 ccatagcggg agaaccggcc aacattgtcg cgctggtcga ggaatacatg gactggctgc
     4981 atcagagccc cgtaccaaaa ttgttgtttt ggggaacgcc aggggtgctg atacctcccg
     5041 cagaggcggc cagactcgcg aaatccctcc ccaattgtaa agccgttgat atagggcccg
     5101 gtttgaatct tttgcaagag gataatccag atttgatagg ttctgagatt gcgagatggt
     5161 tgagcacact ggaaatctca ggctaacgaa ggtaagccta tccctaaccc tctcctcggt
     5221 ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     5281 gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga ggatccagac
     5341 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5401 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5461 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     5521 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagtcgacc
     5581 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     5641 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     5701 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     5761 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     5821 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     5881 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     5941 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     6001 ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7141 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7441 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7681 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7801 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     7861 taggcgtatc acgaggccct ttcgtctt
//
