LOCUS       sequence-313606-        6424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tau in Sf9 cells with a ZZ affinity tag and a ybbR tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6424)
  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 6424)
  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_ybbR_Tau
FEATURES             Location/Qualifiers
     source          1..6424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(520..525,526..542,543..548))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(619..652)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        729..820
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     745..762
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             887..910
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             953..1126
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1127..1300
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             1358..1378
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1400..1420
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2910..3044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2947..2966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3001..3020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3165..3330
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     3337..4042
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3425..3797
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3454..4042)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      4232..4820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4721..4740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     5076..5903
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5076..5200
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5238..5843)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  5932..6155
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6222..6424,1..331))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
       61 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
      121 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
      181 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
      241 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
      301 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
      361 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
      421 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
      481 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
      541 tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
      601 ggcagcagcg aagtcgccat aacttcgtat agcatacatt atacgaagtt atctgtaact
      661 ataacggtcc taaggtagcg agtttaaaca ctagtatcga ttcgcgacct actccggaat
      721 attaatagat catggagata attaaaatga taaccatctc gcaaataaat aagtatttta
      781 ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt attcataccg
      841 tcccaccatc gggcgcggat ctcggtccga aaccatgtcg tactaccatc accatcacca
      901 tcaccatcac ggttctatgt gcatggcagg ccttgcgcaa cacgatgaag ccgtggacaa
      961 caaattcaac aaagaacaac aaaacgcgta ctatgagatc ttacatttac ctaacttaaa
     1021 cgaaggacaa cgaaacgcct tcatccaaag tttaaaagat gacccaagcc aaagcgctaa
     1081 ccttttagca gaagctaaaa agctaaatga tgctcaggcg ccgaaagtag acaacaaatt
     1141 caacaaagaa caacaaaacg cgttctatga gatcttacat ttacctaact taaacgaaga
     1201 acaacgaaac gccttcatcc aaagtttaaa agatgaccca agccaaagcg ctaacctttt
     1261 agcagaagct gaaaagctaa atgatgctca ggcgccgaaa gtagacgcaa attccgcggg
     1321 gaagtcaacc gattatgata ttccaactac tgctagcgag aacctatatt tccagggaga
     1381 ttatgatatc cctacgactg agaatcttta ttttcagggc atggactcat tggagtttat
     1441 cgcatcaaag ctcgcagctg agccccgcca ggagttcgaa gtgatggaag atcacgctgg
     1501 gacgtacggg ttgggggaca ggaaagatca ggggggctac accatgcacc aagaccaaga
     1561 gggtgacacg gacgctggcc tgaaagaatc tcccctgcag acccccactg aggacggatc
     1621 tgaggaaccg ggctctgaaa cctctgatgc taagagcact ccaacagcgg aagatgtgac
     1681 agcaccctta gtggatgagg gagctcccgg caagcaggct gccgcgcagc cccacacgga
     1741 gatcccagaa ggaaccacag ctgaagaagc aggcattgga gacaccccca gcctggaaga
     1801 cgaagctgct ggtcacgtga cccaagctcg catggtcagt aaaagcaaag acgggactgg
     1861 aagcgatgac aaaaaagcca agggggctga tggtaaaacg aagatcgcca caccgcgggg
     1921 agcagcccct ccaggccaga agggccaggc caacgccacc aggattccag caaaaacccc
     1981 gcccgctcca aagacaccac ccagctctgg tgaacctcca aaatcagggg atcgcagcgg
     2041 ctacagcagc cccggctccc caggcactcc cggcagccgc tcccgcaccc cgtcccttcc
     2101 aaccccaccc acccgggagc ccaagaaggt ggcagtggtc cgtactccac ccaagtcgcc
     2161 gtcttccgcc aagagccgcc tgcagacagc ccccgtgccc atgccagacc tgaagaatgt
     2221 caagtccaag atcggctcca ctgagaacct gaagcaccag ccgggaggcg ggaaggtgca
     2281 gataattaat aagaagctgg atcttagcaa cgtccagtcc aagtgtggct caaaggataa
     2341 tatcaaacac gtcccgggag gcggcagtgt gcaaatagtc tacaaaccag ttgacctgag
     2401 caaggtgacc tccaagtgtg gctcattagg caacatccat cataaaccag gaggtggcca
     2461 ggtggaagta aaatctgaga agcttgactt caaggacaga gtccagtcga agattgggtc
     2521 cctggacaat atcacccacg tccctggcgg aggaaataaa aagattgaaa cccacaagct
     2581 gaccttccgc gagaacgcca aagccaagac agaccacggg gcggagatcg tgtacaagtc
     2641 gccagtggtg tctggggaca cgtctccacg gcatctcagc aatgtctcct ccaccggcag
     2701 catcgacatg gtagactcgc cccagctcgc cacgctagct gacgaggtgt ctgcctccct
     2761 ggccaagcag ggtttgtaat gagcttgtcg agaagtacta gaggatcata atcagccata
     2821 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2881 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2941 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3001 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     3061 ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     3121 gagcaataac tatcataacc cctagggtat acccatctaa ttggaaccag ataagtgaaa
     3181 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     3241 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     3301 cccagttccc aactattttg tccgcccaca accggtggag gaaattctcc ttgaagtttc
     3361 cctggtgttc aaagtaaagg agtttgcacc agacgcacct ctgttcactg gtccggcgta
     3421 ttaaaacacg atacattgtt attagtacat ttattaagcg ctagattctg tgcgttgttg
     3481 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
     3541 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa
     3601 atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga
     3661 accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg
     3721 tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac
     3781 gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg
     3841 actcgacgta aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt
     3901 attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga
     3961 ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt
     4021 tgtagttgag ctttttggaa ttaccggttg acttgggtca actgtcagac caagtttact
     4081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4501 ggttggactc aagacgatag ttaccgaata aggcgcagcg gtcgggctga acggggggtt
     4561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattgactt gggtcgctct
     4921 tcctgtggat gcgcaggtat gtacaggaag aggtttatac taaactgtta cattgcaaac
     4981 gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac gcatttctac
     5041 aaccacgact ctaagtgtgt gggtgaagtc atgcatgcat cttttaatca aatcccaaga
     5101 tgtgtataaa ccaccaaact gccaaaaaat gaaaactgtc gacaagctct gtccgtttgc
     5161 tggcaactgc aagggtctca atcctatttg taattattga ataataaaac aattataaat
     5221 gtcaaatttg ttttttatta acgatacaaa ccaaacgcaa caagaacatt tgtagtatta
     5281 tctataattg aaaacgcgta gttataatcg ctgaggtaat atttaaaatc attttcaaat
     5341 gattcacagt taatttgcga caatataatt ttattttcac ataaactaga cgccttgtcg
     5401 tcttcttctt cgtattcctt ctctttttca tttttctctt cataaaaatt aacatagtta
     5461 ttatcgtatc catatatgta tctatcgtat agagtaaatt ttttgttgtc ataaatatat
     5521 atgtcttttt taatggggtg tatagtaccg ctgcgcatag tttttctgta atttacaaca
     5581 gtgctatttt ctggtagttc ttcggagtgt gttgctttaa ttattaaatt tatataatca
     5641 atgaatttgg gatcgtcggt tttgtacaat atgttgccgg catagtacgc agcttcttct
     5701 agttcaatta caccattttt tagcagcacc ggattaacat aactttccaa aatgttgtac
     5761 gaaccgttaa acaaaaacag ttcacctccc ttttctatac tattgtctgc gagcagttgt
     5821 ttgttgttaa aaataacagc cattgtaatg agacgcacaa actaatatca caaactggaa
     5881 atgtctatca atatatagtt gctgattgcg cagatgccct gcgtaagcgg gtgtgggcgg
     5941 acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta
     6001 gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt
     6061 ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag
     6121 aggggcgtgg ccaagggcat gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     6181 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     6241 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     6301 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     6361 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     6421 gcga
//
