LOCUS       sequence-313614-        7348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MAP7 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 7348)
  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 7348)
  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_MAP7
FEATURES             Location/Qualifiers
     source          1..7348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        71..162
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     87..104
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             229..252
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             295..468
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             469..642
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             700..720
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             742..762
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3176..3310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3213..3232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3267..3286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3431..3596
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     3603..4308
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3691..4063
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3720..4308)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      4498..5086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4987..5006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     5342..6169
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5342..5466
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5504..6109)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  6198..6421
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6488..7021)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7210..7215,7216..7232,7233..7238))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(7309..7342)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ctataacggt cctaaggtag cgagtttaaa cactagtatc gattcgcgac ctactccgga
       61 atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt
      121 tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac
      181 cgtcccacca tcgggcgcgg atctcggtcc gaaaccatgt cgtactacca tcaccatcac
      241 catcaccatc acggttctat gtgcatggca ggccttgcgc aacacgatga agccgtggac
      301 aacaaattca acaaagaaca acaaaacgcg tactatgaga tcttacattt acctaactta
      361 aacgaaggac aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct
      421 aaccttttag cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacaacaaa
      481 ttcaacaaag aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa
      541 gaacaacgaa acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt
      601 ttagcagaag ctgaaaagct aaatgatgct caggcgccga aagtagacgc aaattccgcg
      661 gggaagtcaa ccgattatga tattccaact actgctagcg agaacctata tttccaggga
      721 gattatgata tccctacgac tgagaatctt tattttcagg gcatggactc attggagttt
      781 atcgcatcaa agctcgcagc ggagctagga gctggcggcg acggccacag gggcggcgac
      841 ggcgcagtgc gaagcgaaac agcacccgac agctacaaag tgcaagataa gaaaaatgcc
      901 tccagccgcc ctgcctctgc aatttcagga caaaataaca accactcagg aaataaacca
      961 gaccctccgc ctgtgttacg tgttgatgac cggcagcggc tggcccggga gcgacgtgag
     1021 gaacgggaga aacagctagc tgcaagagaa atagtgtggt tagaaagaga agagcgagcc
     1081 aggcagcact acgagaagca cctggaagag cggaagaaga ggttggagga gcagaggcag
     1141 aaggaggagc ggaggagggc tgctgtggag gagaagcgga ggcagagact tgaggaggac
     1201 aaagaacgcc acgaagctgt tgtacggcgc acaatggaaa ggagccagaa gccaaaacag
     1261 aagcataacc gttggtcgtg gggaggctct ctccatggga gccctagcat ccacagtgca
     1321 gatccagaca ggcggtcagt ttccaccatg aatctttcga aatatgttga tcccgtcatt
     1381 agcaagcggc tctcctcttc atctgcaact ttactaaatt ctccagatag agctcgccgc
     1441 ctgcagctca gcccatggga gagcagcgtt gttaacagac tcctgacgcc cacacattcg
     1501 ttcctggcca gaagtaaaag cacagctgcc ttgtctggag aagcagcatc ttgcagcccc
     1561 atcatcatgc cctacaaagc tgcacactct agaaattcga tggatcgacc aaaactcttt
     1621 gtaacaccac ctgagggctc ttctcgcagg aggatcattc atggcacagc gagctataaa
     1681 aaagaaagag agagagaaaa tgtactcttc ctcacatctg gcacccgaag ggctgtatct
     1741 ccatctaatc ccaaagcaag acaaccagct cgctcccgac tttggcttcc gtccaagtct
     1801 cttcctcatt tgcctggcac acccagaccg acatcctcct tgccacccgg ctcagtcaaa
     1861 gctgctcctg ctcaggtccg gcccccatcc cccggcaaca tccgccctgt caagagggaa
     1921 gtcaaagtgg agcctgagaa gaaagatcct gagaaggaac ctcagaaagt tgccaatgag
     1981 ccctcactaa agggcagagc acctttagtg aaggtagaag aagccacagt tgaagagcgg
     2041 acacctgctg aaccagaagt tggccctgct gctccagcca tggccccagc tccagcctcg
     2101 gccccagctc cagcctcggc cccagctcca gccccggtcc ccaccccagc catggtctca
     2161 gccccgtcat ccactgtgaa tgccagtgct tctgttaaga cttctgcagg caccaccgac
     2221 ccagaggagg ccacaaggct tctagctgag aagaggcggc tggcccgaga gcagagagaa
     2281 aaggaagaaa gggagaggag ggagcaggaa gagcttgaaa gacaaaagag agaggaattg
     2341 gctcaacgtg tggctgaaga gaggacgact cgccgtgagg aggagtcgcg caggctggaa
     2401 gccgagcagg cccgggagaa ggaggagcag ctgcagcggc aggcggagga gcgggcgctg
     2461 cgcgagtggg aggaggcaga gcgcgcccag aggcagaaag aagaagaagc tcgcgttcgt
     2521 gaagaagcag agagggtccg gcaggaacga gagaagcatt tccagagaga agagcaagag
     2581 cgcctggaga gaaagaagcg acttgaggag attatgaaaa gaaccaggag aacagaagct
     2641 acagataaga aaaccagtga tcagagaaac ggtgatatag ccaagggagc tctcactgga
     2701 ggaacagagg tgtctgcact tccatgtaca acaaacgctc cgggaaatgg aaagccagtt
     2761 ggtagcccac atgtggttac ctcacaccag tcaaaagtga cagtggagag cactcccgat
     2821 ttggaaaaac aaccaaatga aaatggtgta tctgttcaga atgaaaattt tgaagaaatt
     2881 ataaacttac ccattggatc taaaccatcc agattagatg tcaccaacag tgagagccca
     2941 gaaattcctt tgaatccaat tttggccttt gatgatgaag ggacacttgg gcccctgcct
     3001 caggtagatg gtgttcagac acagcagact gcagaagtta tataatgagc ttgtcgagaa
     3061 gtactagagg ctcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3121 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3301 tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
     3361 aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta gggtataccc
     3421 atctaattgg aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     3481 cgtattaact tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     3541 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacaaccg
     3601 gtggaggaaa ttctccttga agtttccctg gtgttcaaag taaaggagtt tgcaccagac
     3661 gcacctctgt tcactggtcc ggcgtattaa aacacgatac attgttatta gtacatttat
     3721 taagcgctag attctgtgcg ttgttgattt acagacaatt gttgtacgta ttttaataat
     3781 tcattaaatt tataatcttt agggtggtat gttagagcga aaatcaaatg attttcagcg
     3841 tctttatatc tgaatttaaa tattaaatcc tcaatagatt tgtaaaatag gtttcgatta
     3901 gtttcaaaca agggttgttt ttccgaaccg atggctggac tatctaatgg attttcgctc
     3961 aacgccacaa aacttgccaa atcttgtagc agcaatctag ctttgtcgat attcgtttgt
     4021 gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt
     4081 tcatcactgt cgttagtgta caattgactc gacgtaaaca cgttaaatag agcttggaca
     4141 tatttaacat cgggcgtgtt agctttatta ggccgattat cgtcgtcgtc ccaaccctcg
     4201 tcgttagaag ttgcttccga agacgatttt gccatagcca cacgacgcct attaattgtg
     4261 tcggctaaca cgtccgcgat caaatttgta gttgagcttt ttggaattac cggttgactt
     4321 gggtcaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5161 atcccctgat tgacttgggt cgctcttcct gtggatgcgc aggtatgtac aggaagaggt
     5221 ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt
     5281 taatcaaggc tctgacgcat ttctacaacc acgactctaa gtgtgtgggt gaagtcatgc
     5341 atgcatcttt taatcaaatc ccaagatgtg tataaaccac caaactgcca aaaaatgaaa
     5401 actgtcgaca agctctgtcc gtttgctggc aactgcaagg gtctcaatcc tatttgtaat
     5461 tattgaataa taaaacaatt ataaatgtca aatttgtttt ttattaacga tacaaaccaa
     5521 acgcaacaag aacatttgta gtattatcta taattgaaaa cgcgtagtta taatcgctga
     5581 ggtaatattt aaaatcattt tcaaatgatt cacagttaat ttgcgacaat ataattttat
     5641 tttcacataa actagacgcc ttgtcgtctt cttcttcgta ttccttctct ttttcatttt
     5701 tctcttcata aaaattaaca tagttattat cgtatccata tatgtatcta tcgtatagag
     5761 taaatttttt gttgtcataa atatatatgt cttttttaat ggggtgtata gtaccgctgc
     5821 gcatagtttt tctgtaattt acaacagtgc tattttctgg tagttcttcg gagtgtgttg
     5881 ctttaattat taaatttata taatcaatga atttgggatc gtcggttttg tacaatatgt
     5941 tgccggcata gtacgcagct tcttctagtt caattacacc attttttagc agcaccggat
     6001 taacataact ttccaaaatg ttgtacgaac cgttaaacaa aaacagttca cctccctttt
     6061 ctatactatt gtctgcgagc agttgtttgt tgttaaaaat aacagccatt gtaatgagac
     6121 gcacaaacta atatcacaaa ctggaaatgt ctatcaatat atagttgctg attgcgcaga
     6181 tgccctgcgt aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta
     6241 aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt
     6301 atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga
     6361 agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa gggcatgtaa agactatatt
     6421 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     6481 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     6541 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     6601 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     6661 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     6721 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     6781 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     6841 gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc
     6901 gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc
     6961 cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca
     7021 tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
     7081 gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg
     7141 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac
     7201 ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt
     7261 ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgccat aacttcgtat
     7321 agcatacatt atacgaagtt atctgtaa
//
