LOCUS       sequence-468484-       10125 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 10125)
  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 10125)
  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:pCYP86A2:Gus_introns
FEATURES             Location/Qualifiers
     source          1..10125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     223..242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      366..618
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      3937..4189
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4255..4279
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4367..5077
                     /label=oriV
                     /note="incP origin of replication"
     CDS             5372..6163
                     /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      6504..7092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6993..7012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7428..8576
                     /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    8722..8746
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        9248..9593
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     9536..9556
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(9684..10125,1..353)
                     /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(9738..9757)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
       61 ccgaactgtt cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc
      121 acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
      181 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
      241 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
      301 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcttgtc
      361 aagcagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
      421 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
      481 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
      541 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
      601 catctatgtt actagatcga cgctgcaaga attcaagctt ggagtatatc taaggtaaca
      661 catgtatata tatgtcacat agacattact agtatatatt atgtacttct atcatatatt
      721 tatgatattg cagttgcagc gtacacaagt cagctccttt tgacttttac atctcatgaa
      781 tgcattgcca tgacatctaa tcttactcga gatttgtgca tgcacattat tcacttttgt
      841 cttttgcaat ttttgttatt gtaaaaaaag gaaaaacaaa tgtaaaagag agagcgacca
      901 gaaaggtcta actaaaccta aagagtcaat gaaatgtgtt tctttcttgt gggattaatc
      961 aattcactct ttaacacttc tttataccat tgaagaaatt agatgaaaga gtcacgagtt
     1021 gcttaccaaa tccctcacaa gaattgagaa ctgataaacc aaattgagaa gattaaatat
     1081 cacgtctcct tttgatctct attataatta atcgaaaata aaaataagag ttttcaacaa
     1141 aacgtgatca ttggtttacg atcacttgca aagtcaaacc taaaacgtag cattagtaca
     1201 ctaaccttaa atactaatta tatcatgcaa accctaatgt cattacctaa ctatacatgt
     1261 gtaatgtgtt caacagatct tcttaaccca cattagatca atattaaaca ataaaaagat
     1321 tcttatatat tctactactt acttcttctt attcccatcc atatttttct gtgcctttag
     1381 gttctcaact aatctcattt aatttagcta gcacacagag aaacacacac gtatataaat
     1441 aatatgataa cacacaaaaa gactcatata tataaataat tagagtcatt aaatgtggat
     1501 tcatcattaa atgaaacaac tcttcttctc tgtacaattt ctcttcacac cttcaccaaa
     1561 ttctttgact tcaaaaatct tataaaattt atatatctcc aaaaccataa aaccaaaacg
     1621 agttttcaca aataaattac ttagttgaaa tttcaaatct cattcaatta gggtacactc
     1681 tctcaacaat ccacattaat gagggttgct gcatctgatg gctagcagta acagttttat
     1741 cgcctccact tcttaatgcc atctttttcc tcttccctct ccttctctat atatattttc
     1801 tgactctgca aaaccttaat tcatccatct ctcaaacacc atttttggaa acaccatttc
     1861 atattcctta aacttttcca ttttagtatc atttcatatt cattgataat gggtcagtcc
     1921 cttatgttac gtcctgtaga aaccccaacc cgtgaaatca aaaaactcga cggcctgtgg
     1981 gcattcagtc tggatcgcga aaactgtgga attgatcagc gttggtggga aagcgcgtta
     2041 caagaaagcc gggcaattgc tgtgccaggc agttttaacg atcagttcgc cgatgcagat
     2101 attcgtaatt atgcgggcaa cgtctggtat cagcgcgaag tctttatacc gaaaggtaag
     2161 tcttactctc tcttttttgg tctgtatttt taattttttg aagtatacta tttgtactga
     2221 cgctaataat cttttttcag gttgggcagg ccagcgtatc gtgctgcgtt tcgatgcggt
     2281 cactcattac ggcaaagtgt gggtcaataa tcaggaagtg atggagcatc agggcggcta
     2341 tacgccattt gaagccgatg tcacgccgta tgttattgcc gggaaaagtg tacgtatcac
     2401 cgtttgtgtg aacaacgaac tgaactggca gactatcccg ccgggaatgg tgattaccga
     2461 cgaaaacggc aagaaaaagc agtcttactt ccatgatttc tttaactatg ccggaatcca
     2521 tcgcagcgta atgctctaca ccacgccgaa cacctgggtg gacgatatca ccgtggtgac
     2581 gcatgtcgcg caagactgta accacgcgtc tgttgactgg caggtacttc atgcttcaac
     2641 gtgtaactta agagatactg tgtgaaattt tatatttcca tacatttgct tgacctttgc
     2701 tttttgtcaa tttttttccc cttacaggtg gtggccaatg gtgatgtcag cgttgaactg
     2761 cgtgatgcgg atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg
     2821 gtgaatccgc acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc
     2881 aaaagccaga cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg
     2941 aagggcgaac agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat
     3001 gaagatgcgg acttgcgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca
     3061 ttaatggact ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag
     3121 atgctcgact gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc
     3181 tttaacctct ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc
     3241 gaagaggcag tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata
     3301 gcgcgtgaca aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc
     3361 cgtccgcaag gtgcacggga atatttcgcg ccactggcgg aagcaacgcg taaactcgac
     3421 ccgacgcgtc cgatcacctg cgtcaatgta atgttctgcg acgctcacac cgataccatc
     3481 agcgatctct ttgatgtgct gtgcctgaac cgttattacg gatggtatgt ccaaagcggc
     3541 gatttggaaa cggcagagaa ggtactggaa aaagaacttc tggcctggca ggagaaactg
     3601 catcagccga ttatcatcac cgaatacggc gtggatacgt tagccgggct gcactcaatg
     3661 tacaccgaca tgtggagtga agagtatcag tgtgcatggc tggatatgta tcaccgcgtc
     3721 tttgatcgcg tcagcgccgt cgtcggtgaa caggtatgga atttcgccga ttttgcgacc
     3781 tcgcaaggca tattgcgcgt tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa
     3841 ccgaagtcgg cggcttttct gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa
     3901 ccgcagcagg gaggcaaaca atgagcttgt caagcagatc gttcaaacat ttggcaataa
     3961 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     4021 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     4081 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     4141 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg acgctactag
     4201 aggatgcaca tgtgaccgag ggacacgaag tgatccgttt aaactatcag tgtttgacag
     4261 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     4321 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gccagccgcc tttgcgacgc
     4381 tcaccgggct ggttgccctc gccgctgggc tggcggccgt ctatggccct gcaaacgcgc
     4441 cagaaacgcc gtcgaagccg tgtgcgagac accgcggccg ccggcgttgt ggatacctcg
     4501 cggaaaactt ggccctcact gacagatgag gggcggacgt tgacacttga ggggccgact
     4561 cacccggcgc ggcgttgaca gatgaggggc aggctcgatt tcggccggcg acgtggagct
     4621 ggccagcctc gcaaatcggc gaaaacgcct gattttacgc gagtttccca cagatgatgt
     4681 ggacaagcct ggggataagt gccctgcggt attgacactt gaggggcgcg actactgaca
     4741 gatgaggggc gcgatccttg acacttgagg ggcagagtgc tgacagatga ggggcgcacc
     4801 tattgacatt tgaggggctg tccacaggca gaaaatccag catttgcaag ggtttccgcc
     4861 cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt taaaccaata tttataaacc
     4921 ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc
     4981 ccccttctcg aaccctcccg gcccgctaac gcgggcctcc catcccccca ggggctgcgc
     5041 ccctcggccg cgaacggcct caccccaaaa atggcagcgc tggccaattc ccgaggcacg
     5101 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     5161 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     5221 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     5281 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     5341 gggcagtcgc cctaaaacaa agttaaacat catgggggaa gcggtgatcg ccgaagtatc
     5401 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     5461 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagcg atattgattt
     5521 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     5581 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     5641 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     5701 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     5761 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     5821 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     5881 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     5941 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     6001 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     6061 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     6121 ccattacgtg aaaggcgaga tcaccaaggt agtcggcaaa taatgtctag ctagaaattc
     6181 gttcaagccg acgccgcttc gcggcgcggc ttaactcaag cgttagatgc actaagcaca
     6241 taattgctca cagccaaact atcaggtcaa gtctgctttt attattttta agcgtgcata
     6301 ataagcccta cacaaattgg gagatatatc atgctgtcag accaagttta ctcatatata
     6361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcaga tcctagatgt
     7141 ggcgcaacga tgccggcgac aagcaggagc gcaccgactt cttccgcatc aagtgttttg
     7201 gctctcaggc cgaggcccac ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg
     7261 gcaagattcg gaataccaag tacgagaagg acggccagac ggtctacggg accgacttca
     7321 ttgccgataa ggtggattat ctggacacca aggcaccagg cgggtcaaat caggaataag
     7381 ggcacattgc cccggcgtga gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt
     7441 ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc gaggatgccg
     7501 aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag tccgtcggct
     7561 cgatggtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg gctccccctg
     7621 ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tcttgaacag gaggcggcag
     7681 gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag aagcgaaaaa
     7741 ccgccggcga ggacctggca aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac
     7801 acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg
     7861 acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc acgcgcaaca
     7921 agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga
     7981 agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg tggcagcagg
     8041 tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg ttctacgagc
     8101 tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc
     8161 tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg cacctggaat
     8221 cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc
     8281 aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac acgaaattca
     8341 tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac tatttcagct
     8401 cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt
     8461 ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca
     8521 gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa cgctagggcc
     8581 ttgtggggtc agttccggct gggggttcag cagcccctgc tcggatctgt tggaccggac
     8641 agtagtcatg gttgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     8701 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
     8761 acaacttaat aacacattgc ggacgttttt aatgtactgg ggttgaacac tctgtgggtc
     8821 tcatgccgaa ttcggatccg gaggtcaaca tggtggagca cgacactctg gtctactcca
     8881 aaaatgtcaa agatacagtc tcagaagatc aaagggctat tgagactttt caacaaagga
     8941 taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
     9001 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca
     9061 ttcaagatct ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
     9121 tggaaaaaga agaggttcca accacgtcta caaagcaagt ggattgatgt gataacatgg
     9181 tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa
     9241 gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc
     9301 cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc
     9361 atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag
     9421 atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa
     9481 agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc
     9541 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acacgctcga
     9601 gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa
     9661 ttacatttac aattatcgat acaatgattg aacaagatgg attgcacgca ggttctccgg
     9721 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     9781 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     9841 tgtccggtgc cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg ctggccacga
     9901 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     9961 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
    10021 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
    10081 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcgga
//
