LOCUS       sequence-218813-        6566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-Eos-I3 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6566)
  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 6566)
  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-mEos3.2-I3
FEATURES             Location/Qualifiers
     source          1..6566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..108
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     33..50
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             169..186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             208..228
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             241..918
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1502..1636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1539..1558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1593..1612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1746..1911
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2095..2549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2182..2201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2391..2412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2575..2679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2680..2748,2749..3540)
                     /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(2898..2917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3711..4299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4200..4219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4453..4470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4604..4828
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4895..5428)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5598..5603,5604..5620,5621..5626))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(6022..6055)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    6245..6293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6318..6337
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(6429..6538)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 tccggaatat taatagatca tggagataat taaaatgata accatctcgc aaataaataa
       61 gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat
      121 tcataccgtc ccaccatcgg gcgcggatct acaaccatgt cgtactacca tcaccatcac
      181 catcacgatt acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc
      241 atgagtgcga ttaagccaga catgaagatc aaactccgta tggaaggcaa cgtaaacggg
      301 caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg
      361 gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca
      421 ttccattacg gcaacagggt attcgccaaa tatccagaca acatacaaga ctattttaag
      481 cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt
      541 tgcaacgcca gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt
      601 tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag
      661 ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatat tgagatggct
      721 ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag
      781 gagaagggtg tcaagttacc aggcgcccac tttgtggacc actgcattga gattttaagc
      841 catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg
      901 cctgacaatg ccagacgagg tggcggtggc agcatggccg aggcaggggc tgggctgagc
      961 gagaccgtca ctgagacaac ggttaccgtg acaaccgagc ccgagaaccg gagccttacc
     1021 atcaaacttc ggaaacggaa gccagagaaa aaggtagaat ggacaagtga cactgtggac
     1081 aatgaacaca tgggccgccg ctcatccaaa tgctgctgta tttatgagaa acctcgggcc
     1141 tttggcgaga gctccacgga aagtgatgag gaggaagaag agggctgtgg tcatacacac
     1201 tgtgtacgtg gccaccgcaa aggacggcgt cgtgcaaccc taggaccgac ccccaccacc
     1261 cctccccagc ctcctgaccc ttcccagccc cctccagggc caatgcagca ctaatgagtc
     1321 gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt
     1381 cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1441 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1501 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1561 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1621 ttatcatgtc tggatctgat cactgcttga gcctagaaga tccggctgct aacaaagccc
     1681 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actatcataa cccctaggag
     1741 atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     1801 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     1861 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     1921 ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat
     1981 cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct tcgttattaa
     2041 tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc
     2101 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2161 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2221 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2281 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2341 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt
     2401 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2461 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2521 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     2581 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2641 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2701 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2761 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2821 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2881 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2941 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3001 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3061 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3121 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3181 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3241 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3301 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3361 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3421 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3481 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3541 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3601 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4321 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4381 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4441 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4501 ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa
     4561 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa
     4621 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     4681 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     4741 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     4801 ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     4861 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     4921 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     4981 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     5041 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     5101 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     5161 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     5221 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     5281 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     5341 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     5401 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     5461 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     5521 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     5581 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gtttgtgcct
     5641 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat
     5701 ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg
     5761 cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgcccttg cttcaggaga
     5821 tcggtagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc
     5881 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
     5941 ctagtggttg gctacagctt tgtttgtact atcaacaggt tgaactgctg atcaacagat
     6001 cctctacgcg gccgcggtac cataacttcg tatagcatac attatacgaa gttatctggt
     6061 ttaaacgtac ccgtagtggc tatggcaggg cttgccgccc cgacgttggc tgcgagccct
     6121 gggccttcac ccgaacttgg gggttggggt ggggaaaagg aagaaacgcg ggcgtattgg
     6181 tcccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     6241 ttatgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg ccgtcatagc
     6301 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc atctcccggt
     6361 accgcatgct atgcatcagc tgctagcacc atggctcgag atcccgggtg atcaagtctt
     6421 cgtcgagtga ttgtaaataa aatgtaattt acagtatagt attttaatta atatacaaat
     6481 gatttgataa taattcttat ttaactataa tatattgtgt tgggttgaat taaaggtccg
     6541 tatactagta tcgattcgcg acctac
//
