LOCUS       sequence-414437-        7888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Octopus kinesin-1 motor with K282R mutation 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-K282R-HaloTag
FEATURES             Location/Qualifiers
     source          1..7888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..2559
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             4290..5180
                     /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             5188..5229
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5239..5256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5279..5296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5329..5463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5353..5372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5407..5426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6064..6652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6144..6163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6823..7614,7615..7683))
                     /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     7446..7465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7684..7788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7856..7874
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattaattc tatattctaa aaacacaaat gatacttcta aaaaaaaatc atgaatggca
       61 tcaactctga atcaaatctt tgcagatgca cctacttctc atttccactg tcacatcatt
      121 tttccagatc tcgctgcctg ttatgtggcc cacaaaccaa gacacgtttt atggccatta
      181 aagctggctg atcgtcgcca aacaccaaat acataatgaa tatgtacaca ttcgagaaag
      241 aagcgatcaa agaagcgtct tcgggcggag taggagaatg cggaggagaa ggagaacgag
      301 ctgatctagt atctctccac aatccaatgc caactgacca actggccata ttcggagcaa
      361 tttgaagcca atttccatcg cctggcgatc gctccattct tggctatatg tttttcaccg
      421 ttacccgggg ccattttcaa agactcgtcg gcaagataag attgtgtcac tcgctgtctc
      481 tcttcatttg tcgaagaatg ctgaggaatt tcgcgatgac gtcggcgagt attttgaaga
      541 atgagaataa tttgtattta tacgaaaatc agttagtgga attttctaca aaaacatgtt
      601 atctatagat aattttgttg caaaatatgt tgactatgac aaagattgta tgtatatacc
      661 tttaatgtat tctcattttc ttatgtattt ataatggcaa tgatgatact gatgatattt
      721 taagatgatg ccagaccaaa aggcttgaat ttctgcgtct tttgccgaac gcagtgcatg
      781 tgcaattgtt gttttttgga atattcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatgctgcgc
     1081 tctctctagc gccatctcgc ttacgcatgc tcaacgcacc gcatgttgcc gtttcctttt
     1141 atgcgtcatt ttggctcgaa ataggcaatt atttaaacaa agattagtca acgaaaacgc
     1201 taaaataaat aagtctacaa tatggttact tattgccatg tgtgtgcagc caacgatagc
     1261 aacaaaagca acaacacagg tggctttccc tctttcactt tttgtttgca agccgcgtgc
     1321 gagcaagacg gcacgaccgg caaacgcaat tacgctgaca aagagcagac gaagttttgg
     1381 cgaaaaacat caaggcgcct gatacgaatg catttgcaat aacaattgcg atatttaata
     1441 ttgtttatga agctgtttga cttcaaaaca cacaaaaaaa aaaataaaac aaattatttg
     1501 aaagagaatt aggaatcgga cgcttatcgt tagggtaaca acaagaaatg cttactgagt
     1561 cacagcctct ggaaaactgc cgcaagccag agagagagag aaaaagaggg agagcagctt
     1621 agaccgcatg tgcttgtgtg tgaggcgtct ctctcttcgt ctctgttgcg caaacgcata
     1681 gactgcactg aaaaaatcga ttacctattt tttatgaatg aatatttgca ctattactat
     1741 tcaaaactat taagatagca atcacattca atagccaaat actataccac ctgagcgatg
     1801 caacgaaatg atcaatttga gcaaaaatgc tgcatattta ggacggcatc attatagaaa
     1861 tgcttcttgc tgtgtacttt tctctcgtct ggcagctgtt tcgccgttat tgttaaaacc
     1921 ggcttaagtt aggtgtgttt tctacgacta gtgaatgccc tactagaaga tgtgtgttgc
     1981 acaaaatgtc cctggaataa ccaatttgaa gtgcagatag cagtaaacgt aagctaatat
     2041 gaatattatt taactgtaat gttttaatat cgctggacat tactaataaa cccactataa
     2101 acacatgtac atatgtatgt tttggcatac aatgagtagt tggggaaaaa atgtgtaaaa
     2161 gcaccgtgac catcacagca taaagataac cagctgaagt atcgaatatg agtaaccccc
     2221 aaattgaatc acatgccgca actgatagga cccatggaag tacactcttc atggcgatat
     2281 acaagacaca cacaagcacg aacacccagt tgcggaggaa attctccgta aatgaaaacc
     2341 caatcggcga acaattcata cccatatatg gtaaaagttt tgaacgcgac ttgagagcgg
     2401 agagcattgc ggctgataag gttttagcgc taagcgggct ttataaaacg ggctgcggga
     2461 ccagttttca tatcactacc gtttgagttc ttgtgctgtg tggatactcc tcccgacaca
     2521 aagccgctcc atcagccagc agtcgtctaa tccagagacc ccggatcggg gtacctacta
     2581 gtccagtgtg gtggatcaac atggatgcat cttcagaatg caatattaaa gtcatatgcc
     2641 gggtgagacc tctcaatgat gcagaagagc gagcaggctc caagtttata ctcaaatttc
     2701 ctaccgatga ttctataagt atagctggta aagtgttcgt ttttgataaa gtgctcaagc
     2761 caaacgtaag tcaagagtac gtctacaatg ttggcgcaca gccgattgtg gctgatgtcc
     2821 ttagtggata taacgggact attttcgcct acggtcaaac cagttccgga aagacgcaca
     2881 caatggaagg agtccttgat aaaccgtcta tgcacggcat cattccgcgc atagttcagg
     2941 acatattcaa ttatatatac gggatggatg agaacctaga gttccacata aagatttcct
     3001 actatgaaat atacttggat aaaataagag atctcctcga tgtaactaaa acgaacttgt
     3061 cagtgcacga ggacaaaaat cgcgtaccgt atgtcaaggg agccactgag agatttgtca
     3121 gtagcccgga ggaggttatg gaagttattg atgagggtaa aaataaccgg catgtcgctg
     3181 taaccaacat gaacgagcac agttcaaggt ctcactccgt gtttttgatc aacgtaaaac
     3241 aggagaacgt cgaaacgcaa aagaaacttt ccggaaaact gtaccttgtg gaccttgccg
     3301 gatcagaaaa ggtgtctaaa acgggcgcgg aaggagcggt tctcgacgag gcaaaaaaca
     3361 ttaataagag tcttagcgca ctggggaatg ttatcagcgc gctcgctgat gggaacaaga
     3421 gccatgtccc ctaccgagat agtagactga cgcggatctt gcaagaatct cttggaggca
     3481 atgcaaggac aaccatggtc atatgctgta gtcctgcttc ctacaatgaa agcgaaacca
     3541 aatcaacact cctgttcggt caaagggcga aaactatcaa gaatgttgtc agtgtaaacg
     3601 aagaattgac ggccgacgag tggaagcgca gatatgagaa ggaaaaggag aaatgcacga
     3661 aattgaaagc tacgctcgcc agacttgaat ctgaactgca aaaatggcga acaggccaac
     3721 ccgtatctgt tgatgaccag gtggatctca aggctgatat acctgcggaa tcccccgcga
     3781 caagtaccac atccttggct gggaatatgg tggtatcaat gaatgaagga gaccgctcac
     3841 aactggagga ggagcgattg aagctgtatc agcaactcga cgacaaggat gacgagatca
     3901 ataatcagag ccagctcata gagaaattga atgaacaaat gatggagcaa gaagaactca
     3961 tatcccaatc acggaaggat tacgagaatc ttcagcaaga gatgagccga atccagagcg
     4021 acaatgagtc tgctaaggac gaagtgaaag aagtgctgca agcattggag gaattggcga
     4081 tgaactatga ccaaaagtcc caggaagtgg aaaacaagaa taaggagaat gagaatctct
     4141 ccgatgagtt gtcccaaaaa cttagtaccc ttaattcact ccagaatgaa ctggatcaac
     4201 taaaggacag cagtatgcat caccgcaagc gcgtgaccga catgatgatc aaccttctca
     4261 aggacctggg agatggaggt ggaggttcaa tggcggaaat aggcacgggc tttccgtttg
     4321 accctcacta cgttgaggta cttggggaac ggatgcatta cgtagatgta gggccaagag
     4381 acgggacgcc ggtactcttc ctgcacggga atccaacgag cagctacgtg tggagaaata
     4441 tcatcccgca tgtcgccccg actcataggt gtatagctcc agatttgatt ggcatgggaa
     4501 aatcagataa acctgatctt ggttactttt ttgacgacca tgtgaggttt atggacgcat
     4561 tcatagaggc gcttggtctc gaggaggttg tacttgtcat acatgactgg ggttctgcac
     4621 tcggcttcca ctgggcaaag cgcaatcctg aaagggtaaa aggcatcgca tttatggagt
     4681 ttattcggcc tattcccacg tgggatgaat ggccggaatt tgctagagag acgtttcagg
     4741 catttcgcac cacggatgta gggcggaaac ttattattga ccaaaatgta ttcatagagg
     4801 ggactcttcc gatgggtgtt gtgaggccgc ttactgaggt ggaaatggat cactatagag
     4861 agcctttcct taatccagtc gaccgagagc ctctgtggcg gttccccaat gaacttccca
     4921 tagcgggaga accggccaac attgtcgcgc tggtcgagga atacatggac tggctgcatc
     4981 agagccccgt accaaaattg ttgttttggg gaacgccagg ggtgctgata cctcccgcag
     5041 aggcggccag actcgcgaaa tccctcccca attgtaaagc cgttgatata gggcccggtt
     5101 tgaatctttt gcaagaggat aatccagatt tgataggttc tgagattgcg agatggttga
     5161 gcacactgga aatctcaggc taacgaaggt aagcctatcc ctaaccctct cctcggtctc
     5221 gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     5281 tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg
     5341 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5401 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5461 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     5521 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgaccgat
     5581 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     5641 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     5701 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     5761 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     5821 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     5881 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     5941 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     6001 tcagggacag cttcaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6481 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     7141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     7441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7801 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7861 gcgtatcacg aggccctttc gtcttcaa
//
