LOCUS       sequence-313607-        7810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses kinesin-1 (amino acids 1-490) in Sf9 cells with a 
            C-terminal GFP and SNAP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7810)
  AUTHORS   Ferro LS, Fang Q, Eshun-Wilson L, Fernandes J, Jack A, Farrell DP, 
            Golcuk M, Huijben T, Costa K, Gur M, DiMaio F, Nogales E, Yildiz A
  TITLE     Structural and functional insight into regulation of kinesin-1 by 
            microtubule-associated protein MAP7.
  JOURNAL   Science. 2022 Jan 21;375(6578):326-331. doi: 
            10.1126/science.abf6154. Epub 2022 Jan 20.
  PUBMED    35050657
REFERENCE   2  (bases 1 to 7810)
  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: 
            "10.1126/science.abf6154"; journalName: "Science"; date: 
            "2022-01-21- 21"; volume: "375"; issue: "6578"; pages: "326-331"
COMMENT     Sequence Label: pOmnibac_ZZ_linker_K490_GFP_SNAP
FEATURES             Location/Qualifiers
     source          1..7810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(71..104)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        181..272
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     197..214
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             339..362
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             405..578
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             579..752
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             810..830
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             852..872
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2349..3062
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2349..3062
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2379..2407)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3002..3022
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3069..3614
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHLVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    3748..3882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3785..3804)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3839..3858
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4003..4168
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     4175..4880
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           4263..4635
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(4292..4880)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     5914..6741
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5914..6038
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6076..6681)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  6770..6993
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7060..7593)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7782..7787,7788..7804,7805..7810))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
       61 cgaagtcgcc ataacttcgt atagcataca ttatacgaag ttatctgtaa ctataacggt
      121 cctaaggtag cgagtttaaa cactagtatc gattcgcgac ctactccgga atattaatag
      181 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
      241 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      301 tcgggcgcgg atctcggtcc gaaaccatgt cgtactacca tcaccatcac catcaccatc
      361 acggttctat gtgcatggca ggccttgcgc aacacgatga agccgtggac aacaaattca
      421 acaaagaaca acaaaacgcg tactatgaga tcttacattt acctaactta aacgaaggac
      481 aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag
      541 cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag
      601 aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa
      661 acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag
      721 ctgaaaagct aaatgatgct caggcgccga aagtagacgc aaattccgcg gggaagtcaa
      781 ccgattatga tattccaact actgctagcg agaacctata tttccaggga gattatgata
      841 tccctacgac tgagaatctt tattttcagg gcatggcgga cctggccgag tgcaacatca
      901 aagtgatgtg tcgcttcaga cctctcaacg agtctgaagt gaaccgcggc gacaagtaca
      961 tcgccaagtt tcagggagaa gacacggtcg tgatcgcgtc caagccttat gcatttgatc
     1021 gggtgttcca gtcaagcaca tctcaagagc aagtgtataa tgactgtgca aagaagattg
     1081 ttaaagatgt acttgaagga tataatggaa caatatttgc atatggacaa acatcctctg
     1141 ggaagacaca cacaatggag ggtaaacttc atgatccaga aggcatggga attattccaa
     1201 gaatagtgca agatattttt aattatattt actccatgga tgaaaatttg gaatttcata
     1261 ttaaggtttc atattttgaa atatatttgg ataagataag ggacctgtta gatgtttcaa
     1321 agaccaacct ttcagttcat gaagacaaaa accgagttcc ctatgtaaag gggtgcacag
     1381 agcgttttgt atgtagtcca gatgaagtta tggataccat agatgaagga aaatccaaca
     1441 gacatgtagc agttacaaat atgaatgaac atagctctag gagtcacagt atatttctta
     1501 ttaatgtcaa acaagagaac acacaaacgg aacaaaagct gagtggaaaa ctttatctgg
     1561 ttgatttagc tggtagtgaa aaggttagta aaactggagc tgaaggtgct gtgctggatg
     1621 aagctaaaaa catcaacaag tcactttctg ctcttggaaa tgttatttct gctttggctg
     1681 agggtagtac atatgttcca tatcgagata gtaaaatgac aagaatcctt caagattcat
     1741 taggtggcaa ctgtagaacc actattgtaa tttgctgctc tccatcatca tacaatgagt
     1801 ctgaaacaaa atctacactc ttatttggcc aaagggccaa aacaattaag aacacagttt
     1861 gtgtcaatgt ggagttaact gcagaacagt ggaaaaagaa gtatgaaaaa gaaaaagaaa
     1921 aaaataagat cctgcggaac actattcagt ggcttgaaaa tgagctcaac agatggcgta
     1981 atggggagac ggtgcctatt gatgaacagt ttgacaaaga gaaagccaac ttggaagctt
     2041 tcacagtgga taaagatatt actcttacca atgataaacc agcaaccgca attggagtta
     2101 taggaaattt tactgatgct gaaagaagaa agtgtgaaga agaaattgct aaattataca
     2161 aacagcttga tgacaaggat gaagaaatta accagcaaag tcaactggta gagaaactga
     2221 agacgcaaat gttggatcag gaggagcttt tggcatctac cagaagggat caagacaata
     2281 tgcaagctga gctgaatcgc cttcaagcag aaaatgatgc ctctaaagaa gaagtgaaag
     2341 aaggttctat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     2401 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     2461 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     2521 caacacttgt cactactctg acttatggtg ttcaatgctt ttcaagatac ccagatcata
     2581 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     2641 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     2701 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     2761 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
     2821 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     2881 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     2941 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     3001 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     3061 aaggttctat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
     3121 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
     3181 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
     3241 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
     3301 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
     3361 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
     3421 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
     3481 ccgtgcccat tctgatcccc tgccacctgg tggtgcaggg cgacctggac gtggggggct
     3541 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
     3601 agcctgggct gggttaatga gcttgtcgag aagtactaga ggatcataat cagccatacc
     3661 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3721 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3781 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3841 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gcttgagcct
     3901 agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3961 gcaataacta tcataacccc tagggtatac ccatctaatt ggaaccagat aagtgaaatc
     4021 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     4081 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     4141 cagttcccaa ctattttgtc cgcccacaac cggtggagga aattctcctt gaagtttccc
     4201 tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt
     4261 aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat
     4321 ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt
     4381 atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat
     4441 cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac
     4501 cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta
     4561 gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt
     4621 cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac
     4681 tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg ttagctttat
     4741 taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt
     4801 ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg
     4861 tagttgagct ttttggaatt accggttgac ttgggtcaac tgtcagacca agtttactca
     4921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attgacttgg gtcgctcttc
     5761 ctgtggatgc gcaggtatgt acaggaagag gtttatacta aactgttaca ttgcaaacgt
     5821 ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag gctctgacgc atttctacaa
     5881 ccacgactct aagtgtgtgg gtgaagtcat gcatgcatct tttaatcaaa tcccaagatg
     5941 tgtataaacc accaaactgc caaaaaatga aaactgtcga caagctctgt ccgtttgctg
     6001 gcaactgcaa gggtctcaat cctatttgta attattgaat aataaaacaa ttataaatgt
     6061 caaatttgtt ttttattaac gatacaaacc aaacgcaaca agaacatttg tagtattatc
     6121 tataattgaa aacgcgtagt tataatcgct gaggtaatat ttaaaatcat tttcaaatga
     6181 ttcacagtta atttgcgaca atataatttt attttcacat aaactagacg ccttgtcgtc
     6241 ttcttcttcg tattccttct ctttttcatt tttctcttca taaaaattaa catagttatt
     6301 atcgtatcca tatatgtatc tatcgtatag agtaaatttt ttgttgtcat aaatatatat
     6361 gtctttttta atggggtgta tagtaccgct gcgcatagtt tttctgtaat ttacaacagt
     6421 gctattttct ggtagttctt cggagtgtgt tgctttaatt attaaattta tataatcaat
     6481 gaatttggga tcgtcggttt tgtacaatat gttgccggca tagtacgcag cttcttctag
     6541 ttcaattaca ccatttttta gcagcaccgg attaacataa ctttccaaaa tgttgtacga
     6601 accgttaaac aaaaacagtt cacctccctt ttctatacta ttgtctgcga gcagttgttt
     6661 gttgttaaaa ataacagcca ttgtaatgag acgcacaaac taatatcaca aactggaaat
     6721 gtctatcaat atatagttgc tgattgcgca gatgccctgc gtaagcgggt gtgggcggac
     6781 aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga
     6841 cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt
     6901 gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag
     6961 gggcgtggcc aagggcatgt aaagactata ttcgcggcgt tgtgacaatt taccgaacaa
     7021 ctccgcggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc
     7081 gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg
     7141 atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg
     7201 cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc
     7261 gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc
     7321 gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg
     7381 gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc
     7441 gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag
     7501 cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc
     7561 ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc
     7621 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
     7681 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     7741 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
     7801 cttatgtcaa
//
