LOCUS       sequence-440570-        8715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the copper regulated expression of the flippase (FLP) 
            recombinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8715)
  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 8715)
  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: GB4652
FEATURES             Location/Qualifiers
     source          1..8715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             365..385
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1069..1257
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      1870..2122
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2157..2175)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2359..2383
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3684..4313
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4742..5815
                     /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      5881..6075
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6311..6333)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6419..6559
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6474..6493
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6574..6591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6745..7333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6825..6844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7421..8236)
                     /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(7421..8236)
                     /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     8144..8163
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8669..8693
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagagtggt cttgggagag cgatgcgtct tttccgatga accgttccag caaaaaagac
       61 taagcgatgc gtcttttccg ctgaaccgtt ccagcaaaaa agactaagcg atgcgtcttt
      121 tccgctgaac cgttccagca aaaaagacta ggagagcgat gcgtcttttc cgctgaaccg
      181 ttccagcaaa aaagactatc ccgtcctaat acctctacgt gcaagcagta taaaaacaaa
      241 atacataaac ataactaaaa tctcatatcg ttgtaattgt catatactag tggtaattaa
      301 gtccagttac acttaaggtt aaatttgctg actctttttt acctttcatt tctgaaaaat
      361 ggctcctaag aagaagagga aggtgatgag ccagttcgac atcctgtgca agaccccccc
      421 caaggtgctg gtgcggcagt tcgtggagag attcgagagg cccagcggcg agaagatcgc
      481 cagctgtgcc gccgagctga cctacctgtg ctggatgatc acccacaacg gcaccgccat
      541 caagagggcc accttcatga gctacaacac catcatcagc aacagcctga gcttcgacat
      601 cgtgaacaag agcctgcagt tcaagtacaa gacccagaag gccaccatcc tggaggccag
      661 cctgaagaag ctgatccccg cctgggagtt caccatcatc ccttacaacg gccagaagca
      721 ccagagcgac atcaccgaca tcgtgtccag cctgcagctg cagttcgaga gcagcgagga
      781 ggccgacaag ggcaacagcc acagcaagaa gatgctgaag gccctgctgt ccgagggcga
      841 gagcatctgg gagatcaccg agaagatcct gaacagcttc gagtacacca gcaggttcac
      901 caagaccaag accctgtacc agttcctgtt cctggccaca ttcatcaact gcggcaggtt
      961 cagcgacatc aagaacgtgg accccaagag cttcaagctg gtgcagaaca agtacctggg
     1021 cgtgatcatt cagtgcctgg tgaccgagac taagacaagc gtgtccaggt aagtttctgc
     1081 ttctaccttt gatatatata taataattat cattaattag tagtaatata atatttcaaa
     1141 tatttttttc aaaataaaag aatgtagtat atagcaattg cttttctgta gtttataagt
     1201 gtgtatattt taatttataa cttttctaat atatgaccaa aatttgttga tgtgcaggca
     1261 catctacttt ttcagcgcca gaggcaggat cgaccccctg gtgtacctgg acgagttcct
     1321 gaggaacagc gagcccgtgc tgaagagagt gaacaggacc ggcaacagca gcagcaacaa
     1381 gcaggagtac cagctgctga aggacaacct ggtgcgcagc tacaacaagg ccctgaagaa
     1441 gaacgccccc taccccatct tcgctatcaa gaacggccct aagagccaca tcggcaggca
     1501 cctgatgacc agctttctga gcatgaaggg cctgaccgag ctgacaaacg tggtgggcaa
     1561 ctggagcgac aagagggcct ccgccgtggc caggaccacc tacacccacc agatcaccgc
     1621 catccccgac cactacttcg ccctggtgtc caggtactac gcctacgacc ccatcagcaa
     1681 ggagatgatc gccctgaagg acgagactaa ccccatcgag gagtggcagc acatcgagca
     1741 gctgaagggc agcgccgagg gcagcatcag ataccccgcc tggaacggca tcatcagcca
     1801 ggaggtgctg gactacctga gcagctacat caacaggcgg atctgagctt ggaatggatc
     1861 ttcgatcccg atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
     1921 ggtcttgcga cgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
     1981 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
     2041 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     2101 gtgtcatcta tgttactaga tcgggaattg ccaagctaat tcttgaagac gaaagggcct
     2161 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2221 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2281 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat gggaccgact
     2341 cgcgcttgag acgaagcttg acaggatata ttggcgggta aacctaagag aaaagagcgt
     2401 ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt
     2461 atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc aacccctccg
     2521 ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcatctgaaa acgacatgtc
     2581 gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt
     2641 cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat
     2701 gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga
     2761 cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa
     2821 gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc
     2881 tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact
     2941 ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg
     3001 ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga
     3061 gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg
     3121 aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga
     3181 gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg
     3241 ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg
     3301 gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa
     3361 tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc
     3421 attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg
     3481 cacctctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg
     3541 gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt
     3601 gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat
     3661 aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt
     3721 caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg
     3781 ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag
     3841 atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca
     3901 tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt
     3961 ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat
     4021 gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc
     4081 tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg
     4141 ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga
     4201 gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg
     4261 ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg
     4321 aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg
     4381 cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa
     4441 ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa
     4501 gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg
     4561 aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac
     4621 caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa
     4681 gcggctgggt tgcctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg
     4741 cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct
     4801 ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg
     4861 ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc
     4921 ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt
     4981 cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt
     5041 tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga
     5101 cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct
     5161 ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg
     5221 agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg
     5281 agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac
     5341 gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga
     5401 gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta
     5461 catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga
     5521 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta
     5581 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta
     5641 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat
     5701 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat
     5761 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac
     5821 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gactctttcc
     5881 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
     5941 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
     6001 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
     6061 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac
     6121 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
     6181 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc
     6241 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg
     6301 acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt ctgtaagcgg
     6361 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     6421 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
     6481 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6541 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6721 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7381 aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca
     7441 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     7501 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     7561 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     7621 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     7681 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     7741 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata
     7801 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     7861 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     7921 ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     7981 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     8041 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     8101 tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     8161 acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc
     8221 gttgaatatg gctcataaca gaacttatta tttccttcct cttttctaca gtatttaaag
     8281 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     8341 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata
     8401 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt
     8461 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
     8521 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     8581 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     8641 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacataacaa
     8701 gcttcgtctc agtca
//
