LOCUS       sequence-310974-        8370 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 8370)
  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 8370)
  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..8370
                     /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="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4403..5470
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      5536..5730
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5966..5988)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6074..6214
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6129..6148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6229..6246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6400..6988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6480..6499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7076..7891)
                     /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(7076..7891)
                     /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     7799..7818
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8324..8348
                     /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 gatgcctctg 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 accacaacaa aacaaacaac aaacaacatt acaatttact
     1021 attctagtcg aaatggctaa gagatctatt tgggctggtg atgaagataa taagcctaag
     1081 aagagagaaa cttatgctga tgatactgtt ggtagatttc attctggtta ttctgaaact
     1141 aatgaaagag gtaaggttgt tcctgttgct cttgataagt ggagaatttc tactggtgaa
     1201 caatctgttg ctgatgctgt tgctcaactt tttggtggta ctcctgttga aaatgaagaa
     1261 tctacttctg aaaattttat tgatgttttt actgatagac ctaaggttcc tgttattatt
     1321 gaagctgatg gtattcattg ggatatgaag ctttggctta atggtaagct taagcatcat
     1381 tgtgatggtt ttgattttgt ttctcatgct gatgaagaaa tgattggtca accttgtggt
     1441 tgtcctaagc tttttgatga aagaaaggct gctgctaagg aatatgatgc tcctaatcct
     1501 gctattactg ttacttttac tcttgctgat gatcctgaac ttggtagatt taagtttcaa
     1561 actggttctt ggactctttt taaggttctt catgaagctg aagatgatgt tgaaagagtt
     1621 ggtaagggtg gtgctgttct tgctaatctt gaacttgaac ttgttgaata tactcctaag
     1681 agaggtccta tgagaaataa gcttgtttct tattataagc ctactattac tgttcttaag
     1741 tcttataatg atgctattgc tgattaggct tcggccatgc tagagtccgc aaaaatcacc
     1801 agtctctctc tacaaatcta tctctctcta tttttctcca gaataatgtg tgagtagttc
     1861 ccagataagg gaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac
     1921 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     1981 ccaaaatcca gtgacctcgc ttgagacgaa gctttgacag 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 gatccgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     3361 aaccagaaag gcgggtcagg caagacgacc atcggaaccc 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 atagatcgcg cagctaccag
     4201 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     4261 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     4321 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     4381 aagcccgagg aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     4441 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     4501 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     4561 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     4621 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     4681 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     4741 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     4801 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     4861 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     4921 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     4981 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     5041 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     5101 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     5161 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     5221 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     5281 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     5341 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     5401 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     5461 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     5521 aaaaggactc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     5581 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     5641 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     5701 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     5761 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     5821 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     5881 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg
     5941 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggtg acggtcacag
     6001 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6061 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     6121 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6181 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     6241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6841 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcattcta ggtgattaga
     7081 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     7141 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     7201 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     7261 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     7321 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     7381 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     7441 gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     7501 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     7561 ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat gcatcatcag
     7621 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     7681 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     7741 ctggcgcatc gggcttccca tacaatcggt agattgtcgc acctgattgc ccgacattat
     7801 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggccttg
     7861 agcaagacgt ttcccgttga atatggctca taacagaact tattatttcc ttcctctttt
     7921 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     7981 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     8041 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     8101 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     8161 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     8221 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     8281 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     8341 gtgtaaacat aacaagcttc gtctcagtca
//
