LOCUS       sequence-468513-        9762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Evaluating guard cell specific promotors in Brassica.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9762)
  AUTHORS   Nguyen TH, Krasauskas J, Nguyen TB, Noureen A, Smedley M, Christie 
            JM, Harwood W, Blatt MR, Hundleby P
  TITLE     A Promoter Collection for Cell-Targeted Analysis Within the Stomatal
            Complex.
  JOURNAL   Plant Direct. 2025 Feb 12;9(2):e70045. doi: 10.1002/pld3.70045. 
            eCollection 2025 Feb.
  PUBMED    39943924
REFERENCE   2  (bases 1 to 9762)
  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"; journalName: "Plant 
            Direct."; date: "2025-02-12"; volume: "9(2):e70045. doi"; pages: " 
            10.1002/pld3.70045. eCollection 2025 Feb"
COMMENT     Sequence Label: Bo_NPTII:pSNAC1:Gus_introns
FEATURES             Location/Qualifiers
     source          1..9762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    65..89
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      177..887
                     /label=oriV
                     /note="incP origin of replication"
     CDS             1182..1973
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2314..2902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2803..2822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3238..4386
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4532..4556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        5058..5403
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5346..5366
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5494..6288
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5548..5567)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6158..6177
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6301..6553
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      9509..9761
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 acgctactag aggatgcaca tgtgaccgag ggacacgaag tgatccgttt aaactatcag
       61 tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
      121 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gccagccgcc
      181 tttgcgacgc tcaccgggct ggttgccctc gccgctgggc tggcggccgt ctatggccct
      241 gcaaacgcgc cagaaacgcc gtcgaagccg tgtgcgagac accgcggccg ccggcgttgt
      301 ggatacctcg cggaaaactt ggccctcact gacagatgag gggcggacgt tgacacttga
      361 ggggccgact cacccggcgc ggcgttgaca gatgaggggc aggctcgatt tcggccggcg
      421 acgtggagct ggccagcctc gcaaatcggc gaaaacgcct gattttacgc gagtttccca
      481 cagatgatgt ggacaagcct ggggataagt gccctgcggt attgacactt gaggggcgcg
      541 actactgaca gatgaggggc gcgatccttg acacttgagg ggcagagtgc tgacagatga
      601 ggggcgcacc tattgacatt tgaggggctg tccacaggca gaaaatccag catttgcaag
      661 ggtttccgcc cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt taaaccaata
      721 tttataaacc ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg cacgccgaag
      781 gggggtgccc ccccttctcg aaccctcccg gcccgctaac gcgggcctcc catcccccca
      841 ggggctgcgc ccctcggccg cgaacggcct caccccaaaa atggcagcgc tggccaattc
      901 ccgaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
      961 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     1021 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     1081 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     1141 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgggggaa gcggtgatcg
     1201 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     1261 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagcg
     1321 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     1381 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     1441 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     1501 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     1561 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     1621 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     1681 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     1741 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1801 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1861 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     1921 aagaatttgt ccattacgtg aaaggcgaga tcaccaaggt agtcggcaaa taatgtctag
     1981 ctagaaattc gttcaagccg acgccgcttc gcggcgcggc ttaactcaag cgttagatgc
     2041 actaagcaca taattgctca cagccaaact atcaggtcaa gtctgctttt attattttta
     2101 agcgtgcata ataagcccta cacaaattgg gagatatatc atgctgtcag accaagttta
     2161 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2221 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2461 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcaga
     2941 tcctagatgt ggcgcaacga tgccggcgac aagcaggagc gcaccgactt cttccgcatc
     3001 aagtgttttg gctctcaggc cgaggcccac ggcaagtatt tgggcaaggg gtcgctggta
     3061 ttcgtgcagg gcaagattcg gaataccaag tacgagaagg acggccagac ggtctacggg
     3121 accgacttca ttgccgataa ggtggattat ctggacacca aggcaccagg cgggtcaaat
     3181 caggaataag ggcacattgc cccggcgtga gtcggggcaa tcccgcaagg agggtgaatg
     3241 aatcggacgt ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc
     3301 gaggatgccg aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag
     3361 tccgtcggct cgatggtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg
     3421 gctccccctg ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tcttgaacag
     3481 gaggcggcag gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag
     3541 aagcgaaaaa ccgccggcga ggacctggca aaacaggtca gcgaggccaa gcaggccgcg
     3601 ttgctgaaac acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg
     3661 ccgtggccgg acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc
     3721 acgcgcaaca agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac
     3781 aaggacgtga agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg
     3841 tggcagcagg tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg
     3901 ttctacgagc tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc
     3961 gaggaatgcc tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg
     4021 cacctggaat cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg
     4081 tcccgttgcc aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac
     4141 acgaaattca tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac
     4201 tatttcagct cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc
     4261 ggatcggatt ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag
     4321 ttgcgaggca gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa
     4381 cgctagggcc ttgtggggtc agttccggct gggggttcag cagcccctgc tcggatctgt
     4441 tggaccggac agtagtcatg gttgatgggc tgcctgtatc gagtggtgat tttgtgccga
     4501 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
     4561 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggttgaacac
     4621 tctgtgggtc tcatgccgaa ttcggatccg gaggtcaaca tggtggagca cgacactctg
     4681 gtctactcca aaaatgtcaa agatacagtc tcagaagatc aaagggctat tgagactttt
     4741 caacaaagga taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc
     4801 atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga
     4861 aaggctatca ttcaagatct ctctgccgac agtggtccca aagatggacc cccacccacg
     4921 aggagcatcg tggaaaaaga agaggttcca accacgtcta caaagcaagt ggattgatgt
     4981 gataacatgg tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca
     5041 gaagatcaaa gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga
     5101 ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc
     5161 tacaaatgcc atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt
     5221 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc
     5281 acgtctacaa agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa
     5341 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
     5401 acacgctcga gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac
     5461 aacattacaa ttacatttac aattatcgat acaatgattg aacaagatgg attgcacgca
     5521 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5581 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5641 aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg
     5701 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5761 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5821 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5881 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5941 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6001 ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg atctcgtcgt gacccacggc
     6061 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6121 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6181 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6241 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc ttgtcaagca
     6301 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg
     6361 atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc
     6421 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac
     6481 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     6541 atgttactag atcgacgctg caagaattca agcttggagt tactgcaagg tgtctgattt
     6601 ggagtcaaaa tttcggccta gccaaccaac taacttgaaa aatgatttta tattacaaga
     6661 tggagtacta tcatttataa aagttgcctt taagggacaa ttaatattgg acggagggat
     6721 atgagctgtc tttaaaagaa aagatagaat gaaaaacaaa gagcacatga taaaagcacg
     6781 gtaaaaataa atatcagtcg ttttcttctt gacattgttt ttgtttcttt tgaggctgct
     6841 tgccacatac tcccttagta aaaaatactg taatataaaa ggcgtttata gatagctaaa
     6901 gtacggcatg tggtgataaa aaatatatat cagtcgatgc catatatttt ctccaatttt
     6961 gtttcgtggg agatcctgta gagcagctgc tgccacgaca tgcctccgtt tcaaaaaata
     7021 aataaaaaga aaggcaggtg tagacccgag catcagccac gcagaggcgt atccacagtg
     7081 ctcctccgat cccacctatc ggcacgatcc gcggtgtcca cacacgcgca tgccgaccag
     7141 cttcacgccg caccgcacaa cccgcacggc cagctgaagt cggccatgac gcaacccccg
     7201 tgccacccgg ccgcgccttc cccgaaaccc cccacacacg tgcccttttc ctccccaaga
     7261 aacccctcca cgcagccgcc gcccccctac ccccgtccga accttccaga ccactcacgc
     7321 accccccctc ccctatttat ccctccccca tccccccatt taccaccacc atcaccacac
     7381 aacccaactc ctctcactcc ccaacaatct caaacaacag ctcgaggaat caacagcagc
     7441 gactactctg ttcgttgaac caacaatcaa cagacggaca atgggtcagt cccttatgtt
     7501 acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc gacggcctgt gggcattcag
     7561 tctggatcgc gaaaactgtg gaattgatca gcgttggtgg gaaagcgcgt tacaagaaag
     7621 ccgggcaatt gctgtgccag gcagttttaa cgatcagttc gccgatgcag atattcgtaa
     7681 ttatgcgggc aacgtctggt atcagcgcga agtctttata ccgaaaggta agtcttactc
     7741 tctctttttt ggtctgtatt tttaattttt tgaagtatac tatttgtact gacgctaata
     7801 atcttttttc aggttgggca ggccagcgta tcgtgctgcg tttcgatgcg gtcactcatt
     7861 acggcaaagt gtgggtcaat aatcaggaag tgatggagca tcagggcggc tatacgccat
     7921 ttgaagccga tgtcacgccg tatgttattg ccgggaaaag tgtacgtatc accgtttgtg
     7981 tgaacaacga actgaactgg cagactatcc cgccgggaat ggtgattacc gacgaaaacg
     8041 gcaagaaaaa gcagtcttac ttccatgatt tctttaacta tgccggaatc catcgcagcg
     8101 taatgctcta caccacgccg aacacctggg tggacgatat caccgtggtg acgcatgtcg
     8161 cgcaagactg taaccacgcg tctgttgact ggcaggtact tcatgcttca acgtgtaact
     8221 taagagatac tgtgtgaaat tttatatttc catacatttg cttgaccttt gctttttgtc
     8281 aatttttttc cccttacagg tggtggccaa tggtgatgtc agcgttgaac tgcgtgatgc
     8341 ggatcaacag gtggttgcaa ctggacaagg cactagcggg actttgcaag tggtgaatcc
     8401 gcacctctgg caaccgggtg aaggttatct ctatgaactg tgcgtcacag ccaaaagcca
     8461 gacagagtgt gatatctacc cgcttcgcgt cggcatccgg tcagtggcag tgaagggcga
     8521 acagttcctg attaaccaca aaccgttcta ctttactggc tttggtcgtc atgaagatgc
     8581 ggacttgcgt ggcaaaggat tcgataacgt gctgatggtg cacgaccacg cattaatgga
     8641 ctggattggg gccaactcct accgtacctc gcattaccct tacgctgaag agatgctcga
     8701 ctgggcagat gaacatggca tcgtggtgat tgatgaaact gctgctgtcg gctttaacct
     8761 ctctttaggc attggtttcg aagcgggcaa caagccgaaa gaactgtaca gcgaagaggc
     8821 agtcaacggg gaaactcagc aagcgcactt acaggcgatt aaagagctga tagcgcgtga
     8881 caaaaaccac ccaagcgtgg tgatgtggag tattgccaac gaaccggata cccgtccgca
     8941 aggtgcacgg gaatatttcg cgccactggc ggaagcaacg cgtaaactcg acccgacgcg
     9001 tccgatcacc tgcgtcaatg taatgttctg cgacgctcac accgatacca tcagcgatct
     9061 ctttgatgtg ctgtgcctga accgttatta cggatggtat gtccaaagcg gcgatttgga
     9121 aacggcagag aaggtactgg aaaaagaact tctggcctgg caggagaaac tgcatcagcc
     9181 gattatcatc accgaatacg gcgtggatac gttagccggg ctgcactcaa tgtacaccga
     9241 catgtggagt gaagagtatc agtgtgcatg gctggatatg tatcaccgcg tctttgatcg
     9301 cgtcagcgcc gtcgtcggtg aacaggtatg gaatttcgcc gattttgcga cctcgcaagg
     9361 catattgcgc gttggcggta acaagaaagg gatcttcact cgcgaccgca aaccgaagtc
     9421 ggcggctttt ctgctgcaaa aacgctggac tggcatgaac ttcggtgaaa aaccgcagca
     9481 gggaggcaaa caatgagctt gtcaagcaga tcgttcaaac atttggcaat aaagtttctt
     9541 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     9601 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     9661 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     9721 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cg
//
