LOCUS       sequence-218694-        5892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-I3 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5892)
  AUTHORS   Weith M, Seiler J, van den Boom J, Kracht M, Hulsmann J, Primorac I,
            Del Pino Garcia J, Kaschani F, Kaiser M, Musacchio A, Bollen M, 
            Meyer H
  TITLE     Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives
            PP1 Holoenzyme Formation.
  JOURNAL   Mol Cell. 2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020.
  PUBMED    30344098
REFERENCE   2  (bases 1 to 5892)
  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: "Mol Cell. 
            2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020."
COMMENT     Sequence Label: pFL His-I3
FEATURES             Location/Qualifiers
     source          1..5892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..133
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     58..75
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             194..211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             233..253
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    834..968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(871..890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     925..944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1078..1243
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1427..1881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1514..1533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1723..1744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1907..2011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2012..2080,2081..2872)
                     /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(2230..2249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3043..3631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3532..3551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3785..3802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3936..4160
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4227..4760)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4949..4954,4955..4971,4972..4977))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5373..5406)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5596..5644
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5669..5688
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(5780..5889)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atactagtat cgattcgcga cctactccgg aatattaata gatcatggag ataattaaaa
       61 tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt ttgtaataaa
      121 aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg gatctacaac
      181 catgtcgtac taccatcacc atcaccatca cgattacgat atcccaacga ccgaaaacct
      241 gtattttcag ggcgccatgg gatccatggc cgaggcaggg gctgggctga gcgagaccgt
      301 cactgagaca acggttaccg tgacaaccga gcccgagaac cggagcctta ccatcaaact
      361 tcggaaacgg aagccagaga aaaaggtaga atggacaagt gacactgtgg acaatgaaca
      421 catgggccgc cgctcatcca aatgctgctg tatttatgag aaacctcggg cctttggcga
      481 gagctccacg gaaagtgatg aggaggaaga agagggctgt ggtcatacac actgtgtacg
      541 tggccaccgc aaaggacggc gtcgtgcaac cctaggaccg acccccacca cccctcccca
      601 gcctcctgac ccttcccagc cccctccagg gccaatgcag cactaatgag tcgacgagct
      661 cactagtcgc ggccgctttc gaatctagag cctgcagtct cgacaagctt gtcgagaagt
      721 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
      781 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
      841 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
      901 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
      961 tctggatctg atcactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa
     1021 gctgagttgg ctgctgccac cgctgagcaa taactatcat aacccctagg agatccgaac
     1081 cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac
     1141 gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc
     1201 catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc
     1261 ctgttatgtt tttaatcaaa catcctgcca actccatgtg acaaaccgtc atcttcggct
     1321 actttttctc tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt aatgtttgta
     1381 attgactgaa tatcaacgct tatttgcagc ctgaatggcg aatgggacgc gccctgtagc
     1441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1501 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1561 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1621 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1681 acggtttttc gccctttgac gttggagtcc acgttcttaa tagtggactc ttgttccaaa
     1741 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1861 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1921 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1981 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2041 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2101 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2161 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2221 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2281 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2341 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2401 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2461 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2521 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2581 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2641 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2701 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2761 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2821 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2881 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2941 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3001 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3061 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3121 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3181 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3241 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3301 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3361 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3421 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3481 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3541 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3601 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3661 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3721 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3781 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3841 gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac
     3901 ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
     3961 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
     4021 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
     4081 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     4141 gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg
     4201 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     4261 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     4321 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     4381 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     4441 tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca
     4501 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     4561 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     4621 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     4681 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     4741 tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     4801 acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc
     4861 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     4921 gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg
     4981 ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     5041 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg
     5101 tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt
     5161 gcttcaggag atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga
     5221 tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc
     5281 tagctatagt tctagtggtt ggctacagct ttgtttgtac tatcaacagg ttgaactgct
     5341 gatcaacaga tcctctacgc ggccgcggta ccataacttc gtatagcata cattatacga
     5401 agttatctgg tttaaacgta cccgtagtgg ctatggcagg gcttgccgcc ccgacgttgg
     5461 ctgcgagccc tgggccttca cccgaacttg ggggttgggg tggggaaaag gaagaaacgc
     5521 gggcgtattg gtcccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     5581 gaaccccgcg tttatgaaca aacgacccaa cacccgtgcg ttttattctg tctttttatt
     5641 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     5701 catctcccgg taccgcatgc tatgcatcag ctgctagcac catggctcga gatcccgggt
     5761 gatcaagtct tcgtcgagtg attgtaaata aaatgtaatt tacagtatag tattttaatt
     5821 aatatacaaa tgatttgata ataattctta tttaactata atatattgtg ttgggttgaa
     5881 ttaaaggtcc gt
//
