LOCUS       sequence-259174-        7991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 8His-ZZ-LTLT tagged Hook3-SNAPf in insect cells for 
            protein purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7991)
  AUTHORS   Siddiqui N, Zwetsloot AJ, Bachmann A, Roth D, Hussain H, Brandt J, 
            Kaverina I, Straube A
  TITLE     PTPN21 and Hook3 relieve KIF1C autoinhibition and activate 
            intracellular transport.
  JOURNAL   Nat Commun. 2019 Jun 19;10(1):2693. doi: 10.1038/s41467-019-10644-9.
  PUBMED    31217419
REFERENCE   2  (bases 1 to 7991)
  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"; journalName: "Nat 
            Commun."; date: "2019-06-19"; volume: "10(1):2693. doi"; pages: " 
            10.1038/s41467-019-10644-9"
COMMENT     Sequence Label: pFastBacM13-8His-ZZ-LTLT-Hook3-SNAPf
FEATURES             Location/Qualifiers
     source          1..7991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4043..4066
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4514..4534
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4556..4576
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6758..7300
                     /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="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     polyA_signal    7481..7615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7518..7537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7572..7591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7644..7809
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg
     4021 gtccgaaacc atgtcttact atcaccatca ccatcatcac catcacggtt caatgtgcat
     4081 ggctggcctc gcccagcacg acgaagctgt tgacaataag ttcaacaagg aacagcagaa
     4141 cgctttctac gaaatcctcc acctgccaaa cctcaacgaa gaacagcgca acgcattcat
     4201 ccaatcactg aaggacgacc catcccaatc cgcaaacctg ttggccgaag caaagaagct
     4261 gaacgacgct caggctccca aagtggacaa taagttcaat aaagagcagc agaacgcctt
     4321 ctatgagatt ctgcacttgc ctaacctgaa cgaggaacag cgcaatgctt tcatccagtc
     4381 tctgaaggat gatccatcac agagcgctaa tttgctggcc gaggccaaga agctcaatga
     4441 tgctcaagct cccaaggtgg atgctaactc cgccggaaag tccactgact acgacatccc
     4501 tacaactgca agcgagaacc tctacttcca aggcgactat gacatcccaa ctacagagaa
     4561 tctctatttc cagggtgcca tggcgcgccc tatgttcagc gtagagtcgc tggagcgggc
     4621 ggagctgtgc gagagcctcc tcacttggat ccagacattt aatgtggatg caccatgcca
     4681 gaccgtggaa gatttaacga atggggttgt gatggcccag gttcttcaaa agatagatcc
     4741 tgcatatttt gatgaaaatt ggctaaacag aatcaaaact gaagtaggag ataattggag
     4801 gctaaagata agcaatttaa agaaaatttt aaaaggaatc ttggattata atcatgagat
     4861 tttaggacag caaattaatg actttaccct tcctgatgtg aaccttattg gggagcattc
     4921 tgatgcagca gagcttggaa ggatgcttca gctcatctta ggctgtgctg tgaactgtga
     4981 acagaagcaa gagtacatcc aagccattat gatgatggag gaatctgttc aacatgttgt
     5041 catgacagcc attcaagagc tgatgagtaa agaatctcct gtctctgctg gaaatgatgc
     5101 ctatgttgac cttgatcgtc agctgaagaa aactacagag gaactaaatg aagctttgtc
     5161 agcaaaggaa gaaattgctc aaagatgcca tgaactggat atgcaggttg cagcattgca
     5221 ggaagagaaa agtagtttgt tggcagagaa tcaggtatta atggaaagac tcaatcaatc
     5281 tgattctata gaagacccta acagtccagc aggaagaagg catttgcagc tccagactca
     5341 attagaacag ctccaagaag aaacattcag actagaagca gccaaagatg attatcgaat
     5401 acgttgtgaa gagttagaaa aggagatctc tgaacttcgg caacagaatg atgaactgac
     5461 cactttggca gatgaagctc agtctctgaa agatgagatc gacgtgctga gacattcttc
     5521 tgataaagta tctaaactag aaggtcaagt agaatcttat aaaaagaagc tagaagacct
     5581 tggtgattta aggcggcagg ttaaactctt agaagagaag aataccatgt atatgcagaa
     5641 tactgtcagt ctagaggaag agttaagaaa ggccaacgca gcgcgaagtc aacttgaaac
     5701 ctacaagaga caggtagtag aactacaaaa cagattatcc gaagaatcaa agaaagcaga
     5761 taaactagat tttgaatata agcggctaaa agaaaaagtt gacagtcttc aaaaagaaaa
     5821 ggacaggctg agaacagaaa gggattctct gaaggaaacc attgaagagc ttcgttgtgt
     5881 acaagctcaa gaagggcagc tcacaacaca agggttaatg cctcttggaa gtcaggagtc
     5941 ttcagacagt ctagctgcag agattgttac acctgaaatc agggagaaac ttattcgtct
     6001 tcagcatgag aataagatgt taaagcttaa ccaagaaggt tcggacaatg aaaaaatagc
     6061 cttattgcag agccttctag atgatgcaaa tctacgcaag aatgaactgg agacagagaa
     6121 taggctggtg aatcaaagac ttctggaagt acagtcacaa gttgaagaat tacaaaaatc
     6181 tttacaggat caaggctcaa aagcagaaga ttcagtcctt ctaaaaaaga agcttgaaga
     6241 acatctagag aagctgcatg aggccaataa tgaactacag aagaagagag ccattattga
     6301 agatctcgag ccaagattta acaacagctc cttaaaaatt gaagaattac aagaagcttt
     6361 acgaaagaaa gaggaagaaa tgaagcaaat ggaagaacga tacaaaaaat acttagagaa
     6421 agccaaaagt gtcatccgta ctttagatcc taaacagaat caaggagcag caccagaaat
     6481 acaagctctt aaaaatcagc tccaggaacg agaccgactg ttccactcat tagagaaaga
     6541 atatgagaaa acaaagagtc agagagagat ggaagagaaa tatattgtta gtgcctggta
     6601 caatatggga atgaccctgc ataaaaaggc agctgaagat agactggcaa gcacaggctc
     6661 agggcagtca tttctggcga ggcagaggca agcgaccagc agcagaagat catacccagg
     6721 ccacgtgcag ccggccacag caagggatcc accggttgac aaagactgcg aaatgaagcg
     6781 caccaccctg gatagccctc tgggcaagct ggaactgtct gggtgcgaac agggcctgca
     6841 ccgtatcatc ttcctgggca aaggaacatc tgccgccgac gccgtggaag tgcctgcccc
     6901 agccgccgtg ctgggcggac cagagccact gatgcaggcc accgcctggc tcaacgccta
     6961 ctttcaccag cctgaggcca tcgaggagtt ccctgtgcca gccctgcacc acccagtgtt
     7021 ccagcaggag agctttaccc gccaggtgct gtggaaactg ctgaaagtgg tgaagttcgg
     7081 agaggtcatc agctacagcc acctggccgc cctggccggc aatcccgccg ccaccgccgc
     7141 cgtgaaaacc gccctgagcg gaaatcccgt gcccattctg atcccctgcc accgggtggt
     7201 gcagggcgac ctggacgtgg gaggctacga gggcggactc gccgtgaaag agtggctgct
     7261 ggcccacgag ggccacagac tgggcaagcc tggtctgggt taagcggccg ctttcgaatc
     7321 tagagcctgc agtctcgagg catgcggtac caagcttgtc gagaagtact agaggatcat
     7381 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     7441 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     7501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7561 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     7621 actgatatcg cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     7681 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     7741 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     7801 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     7861 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg
     7921 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt tgcagcctga
     7981 atggcgaatg g
//
