LOCUS       sequence-458083-        8942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of the N. nambi NPGA gene under 
            the P35S promoter and the TNOS terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8942)
  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 8942)
  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: GB4829
FEATURES             Location/Qualifiers
     source          1..8942
                     /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"
     terminator      2098..2350
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2385..2403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2591..2615
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3916..4545
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4974..6047
                     /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      6113..6307
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6543..6565)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6651..6791
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6706..6725
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6806..6823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6977..7565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7057..7076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7653..8468)
                     /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(7653..8468)
                     /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     8376..8395
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8901..8925
                     /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 gaaatggtcc aagacacctc aagcgctagt accagtccaa ttttgacaag
     1081 atggtatata gatactagac ctttgactgc ttctaccgca gccttgccac tgttggaaac
     1141 tctgcagcct gcagaccaga taagtgtcca aaagtactac catctgaagg ataagcatat
     1201 gtctctcgca tccaatcttc tcaagtactt gtttgttcac aggaattgcc gaatcccatg
     1261 gtctagcatt gtaatatcaa gaactccaga tccccataga aggccctgtt atataccacc
     1321 ctctggttct caagaggaca gctttaagga tggctacact ggaattaatg tcgagttcaa
     1381 cgtatcccat caagcaagca tggtggcaat agcaggcact gcatttacac ccaactctgg
     1441 aggtgattcc aaacttaagc cagaggttgg catagacatt acttgcgtga atgagcgaca
     1501 ggggcgaaac ggtgaggagc gatccctaga aagtttgagg cagtacatag atattttctc
     1561 agaggttttt tccaccgcag agatggcaaa catcagaaga ttggatgggg tgtcatcaag
     1621 ctccttgagc gccgataggt tggtggacta tggttaccga ttgttttaca cttattgggc
     1681 tcttaaggaa gcatacatca agatgacagg ggaagccctg ctggcaccat ggctcaggga
     1741 gttggaattt agtaatgtcg ttgccccagc agcagtggcc gaatccggag attccgccgg
     1801 agactttgga gagccttata ccggggtacg aacaaccctt tacaaaaact tggtcgagga
     1861 tgttaggata gaggtggctg ctctgggagg tgactacttg ttcgccactg ctgcacgagg
     1921 gggcggtata ggtgcctcca gccgaccagg gggaggccct gatggctcag gaattaggag
     1981 tcaagatccc tggcgaccat ttaaaaagct cgacatagag agagacatac agccttgcgc
     2041 cactggagtc tgtaattgtc tgtctcgagg ttgagcttgg aatggatctt cgatcccgat
     2101 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgacg
     2161 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     2221 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     2281 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     2341 ttactagatc gggaattgcc aagctaattc ttgaagacga aagggcctcg tgatacgcct
     2401 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2461 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2521 gctcatgaga caataaccct gataaatgct tcaataatgg gaccgactcg cgctgtcatg
     2581 agacgaattc tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     2641 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     2701 gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata
     2761 gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt
     2821 cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt
     2881 ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga
     2941 gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca
     3001 accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg
     3061 gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg
     3121 ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg
     3181 ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca
     3241 ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc
     3301 gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga
     3361 agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg
     3421 accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc
     3481 tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg
     3541 ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc
     3601 aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga
     3661 agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc
     3721 aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc
     3781 ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga
     3841 gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat
     3901 acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag
     3961 acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta
     4021 gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg
     4081 ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg
     4141 cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc
     4201 aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc
     4261 gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg
     4321 gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg
     4381 ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca
     4441 ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc
     4501 gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag
     4561 agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca
     4621 aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt
     4681 tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta
     4741 ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg
     4801 agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc
     4861 gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg
     4921 gttgcctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg
     4981 gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga
     5041 agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg
     5101 aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg
     5161 gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga
     5221 tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc
     5281 tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg
     5341 tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga
     5401 tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc
     5461 ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg
     5521 gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg
     5581 ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag
     5641 ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga
     5701 tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga
     5761 cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg
     5821 cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca
     5881 gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa
     5941 atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca
     6001 tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga
     6061 tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggactcttt cctgtggata
     6121 gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc
     6181 caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag
     6241 gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct
     6301 gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt
     6361 cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa
     6421 aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac
     6481 tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa
     6541 aacctctgac acatgcagct cccggtgacg gtcacagctt gtctgtaagc ggatgccggg
     6601 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     6661 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     6721 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6781 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6841 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6901 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6961 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7021 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7081 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7141 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7201 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7261 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7321 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7381 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7441 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7501 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7561 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7621 gttaagggat tttggtcatg cattctaggt gattagaaaa actcatcgag catcaaatga
     7681 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     7741 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     7801 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     7861 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     7921 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     7981 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg
     8041 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     8101 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     8161 cctgggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     8221 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     8281 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     8341 aatcggtaga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     8401 aaatcagcat ccatgttgga atttaatcgc ggccttgagc aagacgtttc ccgttgaata
     8461 tggctcataa cagaacttat tatttccttc ctcttttcta cagtatttaa agatacccca
     8521 agaagctaat tataacaaga cgaactccaa ttcactgttc cttgcattct aaaaccttaa
     8581 ataccagaaa acagcttttt caaagttgtt ttcaaagttg gcgtataaca tagtatcgac
     8641 ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg ctctgtcatc gttacaatca
     8701 acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt
     8761 ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg
     8821 ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg
     8881 gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacataac gaattcgtct
     8941 ca
//
