LOCUS       sequence-313616-        5482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses microtubule-binding domain of tau (amino acids 242-367) 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 5482)
  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 5482)
  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 MTBD
FEATURES             Location/Qualifiers
     source          1..5482
                     /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"
     polyA_signal    1420..1554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1457..1476)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1511..1530
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1675..1840
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     1847..2552
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           1935..2307
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(1964..2552)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      2742..3330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3231..3250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     3586..4413
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             3586..3710
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(3748..4353)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  4442..4665
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4732..5265)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5454..5459,5460..5476,5477..5482))
                     /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 gcatggactc attggagttt atcgcatcaa
      901 agctcgcacg cctgcagaca gcccccgtgc ccatgccaga cctgaagaat gtcaagtcca
      961 agatcggctc cactgagaac ctgaagcacc agccgggagg cgggaaggtg cagataatta
     1021 ataagaagct ggatcttagc aacgtccagt ccaagtgtgg ctcaaaggat aatatcaaac
     1081 acgtcccggg aggcggcagt gtgcaaatag tctacaaacc agttgacctg agcaaggtga
     1141 cctccaagtg tggctcatta ggcaacatcc atcataaacc aggaggtggc caggtggaag
     1201 taaaatctga gaagcttgac ttcaaggaca gagtccagtc gaagattggg tccctggaca
     1261 atatcaccca cgtccctggc ggaggataat gagcttgtcg agaagtacta gaggatcata
     1321 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1381 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1441 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1501 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     1561 ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1621 tgccaccgct gagcaataac tatcataacc cctagggtat acccatctaa ttggaaccag
     1681 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     1741 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     1801 ttccacccct cccagttccc aactattttg tccgcccaca accggtggag gaaattctcc
     1861 ttgaagtttc cctggtgttc aaagtaaagg agtttgcacc agacgcacct ctgttcactg
     1921 gtccggcgta ttaaaacacg atacattgtt attagtacat ttattaagcg ctagattctg
     1981 tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta aatttataat
     2041 ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt
     2101 taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt
     2161 gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg
     2221 ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata
     2281 aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag
     2341 tgtacaattg actcgacgta aacacgttaa atagagcttg gacatattta acatcgggcg
     2401 tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt
     2461 ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg
     2521 cgatcaaatt tgtagttgag ctttttggaa ttaccggttg acttgggtca actgtcagac
     2581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2881 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3001 tgtcttaccg ggttggactc aagacgatag ttaccgaata aggcgcagcg gtcgggctga
     3061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattgactt
     3421 gggtcgctct tcctgtggat gcgcaggtat gtacaggaag aggtttatac taaactgtta
     3481 cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac
     3541 gcatttctac aaccacgact ctaagtgtgt gggtgaagtc atgcatgcat cttttaatca
     3601 aatcccaaga tgtgtataaa ccaccaaact gccaaaaaat gaaaactgtc gacaagctct
     3661 gtccgtttgc tggcaactgc aagggtctca atcctatttg taattattga ataataaaac
     3721 aattataaat gtcaaatttg ttttttatta acgatacaaa ccaaacgcaa caagaacatt
     3781 tgtagtatta tctataattg aaaacgcgta gttataatcg ctgaggtaat atttaaaatc
     3841 attttcaaat gattcacagt taatttgcga caatataatt ttattttcac ataaactaga
     3901 cgccttgtcg tcttcttctt cgtattcctt ctctttttca tttttctctt cataaaaatt
     3961 aacatagtta ttatcgtatc catatatgta tctatcgtat agagtaaatt ttttgttgtc
     4021 ataaatatat atgtcttttt taatggggtg tatagtaccg ctgcgcatag tttttctgta
     4081 atttacaaca gtgctatttt ctggtagttc ttcggagtgt gttgctttaa ttattaaatt
     4141 tatataatca atgaatttgg gatcgtcggt tttgtacaat atgttgccgg catagtacgc
     4201 agcttcttct agttcaatta caccattttt tagcagcacc ggattaacat aactttccaa
     4261 aatgttgtac gaaccgttaa acaaaaacag ttcacctccc ttttctatac tattgtctgc
     4321 gagcagttgt ttgttgttaa aaataacagc cattgtaatg agacgcacaa actaatatca
     4381 caaactggaa atgtctatca atatatagtt gctgattgcg cagatgccct gcgtaagcgg
     4441 gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag
     4501 tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca
     4561 tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt
     4621 cgtattaaag aggggcgtgg ccaagggcat gtaaagacta tattcgcggc gttgtgacaa
     4681 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
     4741 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
     4801 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
     4861 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
     4921 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
     4981 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     5041 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     5101 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     5161 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     5221 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
     5281 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     5341 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     5401 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     5461 gaaccgaaca ggcttatgtc aa
//
