LOCUS       sequence-322025-        8371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of Streptomyces phage 
            PhiC31-encoded recombination directionality factor (RDF) (PhiC31 
            gp3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8371)
  AUTHORS   Bernabe-Orts JM, Quijano-Rubio A, Vazquez-Vilar M, Mancheno-Bonillo 
            J, Moles-Casas V, Selma S, Gianoglio S, Granell A, Orzaez D
  TITLE     A memory switch for plant synthetic biology based on the phage 
            varphiC31 integration system.
  JOURNAL   Nucleic Acids Res. 2020 Apr 6;48(6):3379-3394. doi: 
            10.1093/nar/gkaa104.
  PUBMED    32083668
REFERENCE   2  (bases 1 to 8371)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2020-04-6- 6"; volume: "48"; issue: "6";
            pages: "3379-3394"
COMMENT     Sequence Label: pEGB3 alpha2 P35S:RDF:T35S (GB1508)
FEATURES             Location/Qualifiers
     source          1..8371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2015..2039
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3340..3969
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4398..5471
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5537..5731
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5967..5989)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6075..6215
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6130..6149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6230..6247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6401..6989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6481..6500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7077..7892)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7077..7892)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7800..7819
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8325..8349
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatggcta agagatctat ttgggctggt gatgaagata ataagcctaa
     1081 gaagagagaa acttatgctg atgatactgt tggtagattt cattctggtt attctgaaac
     1141 taatgaaaga ggtaaggttg ttcctgttgc tcttgataag tggagaattt ctactggtga
     1201 acaatctgtt gctgatgctg ttgctcaact ttttggtggt actcctgttg aaaatgaaga
     1261 atctacttct gaaaatttta ttgatgtttt tactgataga cctaaggttc ctgttattat
     1321 tgaagctgat ggtattcatt gggatatgaa gctttggctt aatggtaagc ttaagcatca
     1381 ttgtgatggt tttgattttg tttctcatgc tgatgaagaa atgattggtc aaccttgtgg
     1441 ttgtcctaag ctttttgatg aaagaaaggc tgctgctaag gaatatgatg ctcctaatcc
     1501 tgctattact gttactttta ctcttgctga tgatcctgaa cttggtagat ttaagtttca
     1561 aactggttct tggactcttt ttaaggttct tcatgaagct gaagatgatg ttgaaagagt
     1621 tggtaagggt ggtgctgttc ttgctaatct tgaacttgaa cttgttgaat atactcctaa
     1681 gagaggtcct atgagaaata agcttgtttc ttattataag cctactatta ctgttcttaa
     1741 gtcttataat gatgctattg ctgattaggc ttcggccatg ctagagtccg caaaaatcac
     1801 cagtctctct ctacaaatct atctctctct atttttctcc agaataatgt gtgagtagtt
     1861 cccagataag ggaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa
     1921 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     1981 accaaaatcc agtgacctcg cttgagacga agcttgacag gatatattgg cgggtaaacc
     2041 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     2101 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
     2161 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat
     2221 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
     2281 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
     2341 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     2401 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     2461 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     2521 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     2581 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     2641 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     2701 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     2761 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     2821 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     2881 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     2941 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     3001 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     3061 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     3121 gttcgagccg cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     3181 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     3241 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     3301 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     3361 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     3421 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     3481 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     3541 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     3601 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     3661 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     3721 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     3781 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     3841 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     3901 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     3961 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     4021 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     4081 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     4141 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     4201 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     4261 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     4321 ggttggccag gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc
     4381 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
     4441 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
     4501 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
     4561 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
     4621 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
     4681 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
     4741 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
     4801 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
     4861 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
     4921 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
     4981 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
     5041 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
     5101 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
     5161 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
     5221 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
     5281 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
     5341 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
     5401 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
     5461 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
     5521 gaaaaggact ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
     5581 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
     5641 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
     5701 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
     5761 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
     5821 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
     5881 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc
     5941 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccggt gacggtcaca
     6001 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6061 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     6121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     6181 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     6241 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6301 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6421 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6481 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6541 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6841 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6901 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7021 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtgattag
     7081 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     7141 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     7201 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     7261 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     7321 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     7381 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     7441 tgagcgagtc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     7501 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     7561 tctaatacct ggaatgctgt tttccctggg atcgcagtgg tgagtaacca tgcatcatca
     7621 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     7681 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     7741 tctggcgcat cgggcttccc atacaatcgg tagattgtcg cacctgattg cccgacatta
     7801 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt
     7861 gagcaagacg tttcccgttg aatatggctc ataacagaac ttattatttc cttcctcttt
     7921 tctacagtat ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact
     7981 gttccttgca ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa
     8041 gttggcgtat aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc
     8101 aacgctctgt catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa
     8161 cccggcagct tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc
     8221 cttacaacgg ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt
     8281 gattttgtgc cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt
     8341 ggtgtaaaca taacaagctt cgtctcagtc a
//
