LOCUS       sequence-458088-        8653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of the flippase (FLP) recombinase
            with a nopaline synthase (Nos) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8653)
  AUTHORS   Calvache C, Vazquez-Vilar M, Moreno-Gimenez E, Orzaez D
  TITLE     A quantitative autonomous bioluminescence reporter system with a 
            wide dynamic range for Plant Synthetic Biology.
  JOURNAL   Plant Biotechnol J. 2024 Jan;22(1):37-47. doi: 10.1111/pbi.14146. 
            Epub 2023 Oct 26.
  PUBMED    37882352
REFERENCE   2  (bases 1 to 8653)
  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: "10.1111/pbi.14146";
            journalName: "Plant Biotechnol J"; date: "2024-01"; volume: "22"; 
            issue: "1"; pages: "37-47"
COMMENT     Sequence Label: GB4653
FEATURES             Location/Qualifiers
     source          1..8653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        83..262
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             303..323
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1007..1195
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      1808..2060
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2095..2113)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2297..2321
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3622..4251
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4680..5753
                     /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      5819..6013
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6249..6271)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6357..6497
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6412..6431
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6512..6529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6683..7271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6763..6782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7359..8174)
                     /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(7359..8174)
                     /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     8082..8101
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8607..8631
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagatacat gagaattaag ggagtcacgg tatgaccccc gccgatgacg cgggacaagc
       61 cgttttacgt ttggaactga cagaaccgca acgttgaagg agccactgag ccgcgggttt
      121 ctggagttta atgagctaag cacatacgtc agaaaccatt attgcgcgtt caaaagtcgc
      181 ctaaggtcac tatcagctag caaatatttc ttgtcaaaaa tgctccactg acgttccata
      241 aattcccctc ggtatccaat tagagtctca tattcactct caactcgatc gaggcaatgg
      301 ctcctaagaa gaagaggaag gtgatgagcc agttcgacat cctgtgcaag acccccccca
      361 aggtgctggt gcggcagttc gtggagagat tcgagaggcc cagcggcgag aagatcgcca
      421 gctgtgccgc cgagctgacc tacctgtgct ggatgatcac ccacaacggc accgccatca
      481 agagggccac cttcatgagc tacaacacca tcatcagcaa cagcctgagc ttcgacatcg
      541 tgaacaagag cctgcagttc aagtacaaga cccagaaggc caccatcctg gaggccagcc
      601 tgaagaagct gatccccgcc tgggagttca ccatcatccc ttacaacggc cagaagcacc
      661 agagcgacat caccgacatc gtgtccagcc tgcagctgca gttcgagagc agcgaggagg
      721 ccgacaaggg caacagccac agcaagaaga tgctgaaggc cctgctgtcc gagggcgaga
      781 gcatctggga gatcaccgag aagatcctga acagcttcga gtacaccagc aggttcacca
      841 agaccaagac cctgtaccag ttcctgttcc tggccacatt catcaactgc ggcaggttca
      901 gcgacatcaa gaacgtggac cccaagagct tcaagctggt gcagaacaag tacctgggcg
      961 tgatcattca gtgcctggtg accgagacta agacaagcgt gtccaggtaa gtttctgctt
     1021 ctacctttga tatatatata ataattatca ttaattagta gtaatataat atttcaaata
     1081 tttttttcaa aataaaagaa tgtagtatat agcaattgct tttctgtagt ttataagtgt
     1141 gtatatttta atttataact tttctaatat atgaccaaaa tttgttgatg tgcaggcaca
     1201 tctacttttt cagcgccaga ggcaggatcg accccctggt gtacctggac gagttcctga
     1261 ggaacagcga gcccgtgctg aagagagtga acaggaccgg caacagcagc agcaacaagc
     1321 aggagtacca gctgctgaag gacaacctgg tgcgcagcta caacaaggcc ctgaagaaga
     1381 acgcccccta ccccatcttc gctatcaaga acggccctaa gagccacatc ggcaggcacc
     1441 tgatgaccag ctttctgagc atgaagggcc tgaccgagct gacaaacgtg gtgggcaact
     1501 ggagcgacaa gagggcctcc gccgtggcca ggaccaccta cacccaccag atcaccgcca
     1561 tccccgacca ctacttcgcc ctggtgtcca ggtactacgc ctacgacccc atcagcaagg
     1621 agatgatcgc cctgaaggac gagactaacc ccatcgagga gtggcagcac atcgagcagc
     1681 tgaagggcag cgccgagggc agcatcagat accccgcctg gaacggcatc atcagccagg
     1741 aggtgctgga ctacctgagc agctacatca acaggcggat ctgagcttgg aatggatctt
     1801 cgatcccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     1861 tcttgcgacg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     1921 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     1981 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     2041 gtcatctatg ttactagatc gggaattgcc aagctaattc ttgaagacga aagggcctcg
     2101 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatgg gaccgactcg
     2281 cgcttgagac gaagcttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
     2341 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat
     2401 gtgcatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa cccctccgct
     2461 gctatagtgc agtcggcttc tgacgttcag tgcagccgtc atctgaaaac gacatgtcgc
     2521 acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt tttcttgtcg
     2581 cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat tacgccatga
     2641 acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac gaccaggact
     2701 tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt tccgagaaga
     2761 tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac ctacgccctg
     2821 gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc gacctactgg
     2881 acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca gagccgtggg
     2941 ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc attgccgagt
     3001 tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag
     3061 gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac gcccgcgagc
     3121 tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc gtgcatcgct
     3181 cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag gccaggcggc
     3241 gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc gccgagaatg
     3301 aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac cgtttttcat
     3361 taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca
     3421 cctctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc
     3481 ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt
     3541 atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa
     3601 acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca
     3661 ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt
     3721 ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat
     3781 caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc
     3841 ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc
     3901 gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg
     3961 gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta
     4021 caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc
     4081 gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc
     4141 tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct
     4201 gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag
     4261 gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca
     4321 gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact
     4381 ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga
     4441 ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg agcaaatgaa
     4501 taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca
     4561 ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc
     4621 ggctgggttg cctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg
     4681 tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg
     4741 tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca gaagcacgcc
     4801 ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg caaccgccgg
     4861 cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca gattttttcg
     4921 ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac gtggccgttt
     4981 tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag cttccagacg
     5041 ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat tacgacctgg
     5101 tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa gggaagggag
     5161 acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc tgccggcgag
     5221 ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta aacaccacgc
     5281 acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg gtatccgagg
     5341 gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg ccggagtaca
     5401 tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag aacccggacg
     5461 tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt tttctctacc
     5521 gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg
     5581 aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg
     5641 ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc
     5701 tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg
     5761 agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaagga ctctttcctg
     5821 tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac ccgtacattg
     5881 ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat ataaaagaga
     5941 aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt aaaacccgcc
     6001 tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa gcgcctaccc
     6061 ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc
     6121 gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa gccgcgccgt
     6181 cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt cggtgatgac
     6241 ggtgaaaacc tctgacacat gcagctcccg gtgacggtca cagcttgtct gtaagcggat
     6301 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     6361 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     6421 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     6481 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7321 actcacgtta agggattttg gtcatgcatt ctaggtgatt agaaaaactc atcgagcatc
     7381 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     7441 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     7501 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     7561 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     7621 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     7681 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag tcgaaatacg
     7741 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     7801 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     7861 gttttccctg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     7921 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     7981 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     8041 ccatacaatc ggtagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     8101 ccatataaat cagcatccat gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt
     8161 tgaatatggc tcataacaga acttattatt tccttcctct tttctacagt atttaaagat
     8221 accccaagaa gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa
     8281 ccttaaatac cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt
     8341 atcgacggag ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta
     8401 caatcaacat gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc
     8461 gttcttccga atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg
     8521 ctgacgccgt cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc
     8581 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa cataacaagc
     8641 ttcgtctcag tca
//
