LOCUS       sequence-465273-        5812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of His6-tagged Pseudomonas syringae pv tomato
            type III effector HopF2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5812)
  AUTHORS   Simranjit Kaur, Thomas Colby, Domenika Thieme, Carsten Proksch, 
            Susanne Matschi, eIvan Matic,, Lennart Wirthmueller
  TITLE     Untargeted proteomics identifies plant substrates of the 
            bacterial-derived ADP-ribosyltransferase AvrRpm1
  JOURNAL   bioRxiv 2023.09.25.558804
REFERENCE   2  (bases 1 to 5812)
  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: "bioRxiv 
            2023.09.25.558804"
COMMENT     Sequence Label: pOPINF HopF2
FEATURES             Location/Qualifiers
     source          1..5812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             181..305
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(343..948)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1140..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1449..1727
                     /label=chicken beta-actin promoter
     primer_bind     2006..2028
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2090..2109
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2151..2170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2151..2169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2170..2194
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        2210..2319
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2340..2357
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2367..2390
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3107..3126)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3172..3227
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3226..3245)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3315..3362
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3375..4080
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3463..3835
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3492..4080)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4089..4122)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4290..4877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4369..4388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4890..5681,5682..5750))
                     /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     5513..5532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
       61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
      121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
      181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
      241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
      301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
      361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
      421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
      481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
      541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
      601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
      661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
      721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
      781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
      841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
      901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
      961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccg
     1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct
     1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     1561 gggggggggg cggcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     1621 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1681 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1741 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1801 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctccttcg ggctgtaatt
     1861 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1921 tccgggaggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg ggacggctgc
     1981 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     2041 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     2101 gttattgtgc tgtctcatca ttttggcaaa gaattggatc ggaccgaaat taatacgact
     2161 cactataggg gaattgtgag cggataacaa ttccccggag ttaatccggg acctttaatt
     2221 caacccaaca caatatatta tagttaaata agaattatta tcaaatcatt tgtatattaa
     2281 ttaaaatact atactgtaaa ttacatttta tttacaatca aaggagatat accatggcac
     2341 accatcacca ccatcacagc agcggtctgg aagttctgtt tcagggcccg ggtaatattt
     2401 gcggcacctc gggctcacgt catgtgtata gcccatccca tacacaacga ataacttcag
     2461 ctccctctac atccactcat gttggtggag atacactgac atccattcat cagctttcgc
     2521 atagtcagag agagcagttt ctgaacatgc atgatccaat gagagtaatg ggacttgacc
     2581 atgataccga gcttttcaga acgacggata gtcgctatat aaaaaacgat aaactcgcgg
     2641 gcaatccaca atccatggcg agtatcctta tgcatgaaga actgcgcccc aatcgttttg
     2701 ccagccatac aggtgcccaa ccacacgaag caagggcgta cgttccgaaa agaataaaag
     2761 ccaccgatct aggagttcca tcactgaacg taatgactgg ctcgctagcg cgagacggaa
     2821 ttagagctta tgatcacatg agtgataatc aggtatctgt caaaatgcga ctgggagatt
     2881 ttctcgaaag gggtggcaag gtctatgccg acgcttcgtc tgtagctgac gatggggaaa
     2941 catcacaagc tctgattgtc acattgccca aaggacagaa agtgccggtc gaaagggtct
     3001 gaagctttct agaccagttt gtgattaacc tcaggtgcag gctgcctatc agaaggtggt
     3061 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatcgatctt tttccctctg
     3121 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3181 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3241 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3301 atgcccatat gtaactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3361 tgctgaaagc atgcggagga aattctcctt gaagtttccc tggtgttcaa agtaaaggag
     3421 tttgcaccag acgcacctct gttcactggt ccggcgtatt aaaacacgat acattgttat
     3481 tagtacattt attaagcgct agattctgtg cgttgttgat ttacagacaa ttgttgtacg
     3541 tattttaata attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa
     3601 tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat
     3661 aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat
     3721 ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg
     3781 atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct
     3841 tttgtatttc tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat
     3901 agagcttgga catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg
     3961 tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc
     4021 ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttggaatt
     4081 gcgatcgcat aacttcgtat agcatacatt atacgaagtt ataagctcgg aacgctgcgc
     4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccccct gacgagcatc
     4321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4861 gaaaaaaagg atctcaagaa gatcctttgt taccaatgct taatcagtga ggcacctatc
     4921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4981 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5041 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5101 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5401 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5761 tttcaatatt attgaagcat ttatcagggt tattgtctca tgtccgcgcg tt
//
