LOCUS       sequence-468514-        9833 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 9833)
  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 9833)
  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:pGC1:Gus_introns
FEATURES             Location/Qualifiers
     source          1..9833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             972..1763
                     /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      2104..2692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2593..2612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3028..4176
                     /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    4322..4346
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4848..5193
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5136..5156
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5284..6078
                     /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(5338..5357)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5948..5967
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6091..6343
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      9370..9622
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    9688..9712
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      join(9800..9833,1..677)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gccgctgggc tggcggccgt ctatggccct gcaaacgcgc cagaaacgcc gtcgaagccg
       61 tgtgcgagac accgcggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact
      121 gacagatgag gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca
      181 gatgaggggc aggctcgatt tcggccggcg acgtggagct ggccagcctc gcaaatcggc
      241 gaaaacgcct gattttacgc gagtttccca cagatgatgt ggacaagcct ggggataagt
      301 gccctgcggt attgacactt gaggggcgcg actactgaca gatgaggggc gcgatccttg
      361 acacttgagg ggcagagtgc tgacagatga ggggcgcacc tattgacatt tgaggggctg
      421 tccacaggca gaaaatccag catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa
      481 cctgtctttt aacctgcttt taaaccaata tttataaacc ttgtttttaa ccagggctgc
      541 gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc ccccttctcg aaccctcccg
      601 gcccgctaac gcgggcctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct
      661 caccccaaaa atggcagcgc tggccaattc ccgaggcacg aacccagtgg acataagcct
      721 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
      781 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
      841 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
      901 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
      961 agttaaacat catgggggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     1021 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     1081 cagtggatgg cggcctgaag ccacacagcg atattgattt gctggttacg gtgaccgtaa
     1141 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     1201 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     1261 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     1321 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     1381 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     1441 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     1501 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     1561 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     1621 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     1681 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccattacgtg aaaggcgaga
     1741 tcaccaaggt agtcggcaaa taatgtctag ctagaaattc gttcaagccg acgccgcttc
     1801 gcggcgcggc ttaactcaag cgttagatgc actaagcaca taattgctca cagccaaact
     1861 atcaggtcaa gtctgctttt attattttta agcgtgcata ataagcccta cacaaattgg
     1921 gagatatatc atgctgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1981 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2041 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2101 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2161 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2221 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2281 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2341 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2401 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2461 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2521 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2581 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2641 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2701 caacgcggcc tttttacggt tcctggcaga tcctagatgt ggcgcaacga tgccggcgac
     2761 aagcaggagc gcaccgactt cttccgcatc aagtgttttg gctctcaggc cgaggcccac
     2821 ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg gcaagattcg gaataccaag
     2881 tacgagaagg acggccagac ggtctacggg accgacttca ttgccgataa ggtggattat
     2941 ctggacacca aggcaccagg cgggtcaaat caggaataag ggcacattgc cccggcgtga
     3001 gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt ttgaccggaa ggcatacagg
     3061 caagaactga tcgacgcggg gttttccgcc gaggatgccg aaaccatcgc aagccgcacc
     3121 gtcatgcgtg cgccccgcga aaccttccag tccgtcggct cgatggtcca gcaagctacg
     3181 gccaagatcg agcgcgacag cgtgcaactg gctccccctg ccctgcccgc gccatcggcc
     3241 gccgtggagc gttcgcgtcg tcttgaacag gaggcggcag gtttggcgaa gtcgatgacc
     3301 atcgacacgc gaggaactat gacgaccaag aagcgaaaaa ccgccggcga ggacctggca
     3361 aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac acacgaagca gcagatcaag
     3421 gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg acacgatgcg agcgatgcca
     3481 aacgacacgg cccgctctgc cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg
     3541 ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga agatcaccta caccggcgtc
     3601 gagctgcggg ccgacgatga cgaactggtg tggcagcagg tgttggagta cgcgaagcgc
     3661 acccctatcg gcgagccgat caccttcacg ttctacgagc tttgccagga cctgggctgg
     3721 tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc tgtcgcgcct acaggcgacg
     3781 gcgatgggct tcacgtccga ccgcgttggg cacctggaat cggtgtcgct gctgcaccgc
     3841 ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc aggtcctgat cgacgaggaa
     3901 atcgtcgtgc tgtttgctgg cgaccactac acgaaattca tatgggagaa gtaccgcaag
     3961 ctgtcgccga cggcccgacg gatgttcgac tatttcagct cgcaccggga gccgtacccg
     4021 ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt ccacccgcgt gaagaagtgg
     4081 cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca gcggcctggt ggaacacgcc
     4141 tgggtcaatg atgacctggt gcattgcaaa cgctagggcc ttgtggggtc agttccggct
     4201 gggggttcag cagcccctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc
     4261 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     4321 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     4381 ggacgttttt aatgtactgg ggttgaacac tctgtgggtc tcatgccgaa ttcggatccg
     4441 gaggtcaaca tggtggagca cgacactctg gtctactcca aaaatgtcaa agatacagtc
     4501 tcagaagatc aaagggctat tgagactttt caacaaagga taatttcggg aaacctcctc
     4561 ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc
     4621 tcctacaaat gccatcattg cgataaagga aaggctatca ttcaagatct ctctgccgac
     4681 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agaggttcca
     4741 accacgtcta caaagcaagt ggattgatgt gataacatgg tggagcacga cactctggtc
     4801 tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctattga gacttttcaa
     4861 caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc
     4921 gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
     4981 gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
     5041 agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa agcaagtgga ttgatgtgac
     5101 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     5161 atataaggaa gttcatttca tttggagagg acacgctcga gtataagagc tcatttttac
     5221 aacaattacc aacaacaaca aacaacaaac aacattacaa ttacatttac aattatcgat
     5281 acaatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5341 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5401 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5461 ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5521 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5581 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5641 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5701 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5761 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc
     5821 gacggcgagg atctcgtcgt gacccacggc gatgcctgct tgccgaatat catggtggaa
     5881 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5941 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6001 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6061 cttgacgagt tcttctgagc ttgtcaagca gatcgttcaa acatttggca ataaagtttc
     6121 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
     6181 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
     6241 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     6301 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgacgctg caagaattca
     6361 agcttggaga aacaagaaaa aaaacctata caatcaatct ttaggaattg acgatgtaga
     6421 attgtagatg ataaattttc tcaaatatag atgggcctaa tgaagggtgc cgcttattgg
     6481 atctgaccca ttttgaggac attaatattt tcattggtta taagcctttt aatcaaaatt
     6541 gtcattaaat tgatgtctcc ctctcgggtc attttccttt ctccctcaca attaatgtag
     6601 actttagcaa tttgcacgct gtgctttgtc tttatattta gtaacacaaa cattttgact
     6661 tgtcttgtag agtttttctc ttttattttt ctatccaata tgaaaactaa aagtgttctc
     6721 gtatacatat attaaaatta aagaaaccta tgaaaacacc aatacaaatg cgatattgtt
     6781 ttcagttcga cgtttcatgt ttgttagaaa atttctaatg acgtttgtat aaaatagaca
     6841 attaaacgcc aaacactaca tctgtgtttt cgaacaatat tgcgtctgcg tttccttcat
     6901 ctatctctct cagtgtcaca atgtctgaac taagagacag ctgtaaacta tcattaagac
     6961 ataaactacc aaagtatcaa gctaatgtaa aaattactct catttccacg taacaaattg
     7021 agttagctta agatattagt gaaactaggt ttgaattttc ttcttcttct tccatgcatc
     7081 ctccgaaaaa agggaaccaa tcaaaactgt ttgcatatca aactccaaca ctttacagca
     7141 aatgcaatct ataatctgtg atttatccaa taaaaacctg tgatttatgt ttggctccag
     7201 cgatgaaagt ctatgcatgt gatctctatc caacatgagt aattgttcag aaaataaaaa
     7261 gtagctgaaa tgtatctata taaagaatca tccacaagta ctattttcac acactacttc
     7321 aaaatcacta ctcaagaaat aatgggtcag tcccttatgt tacgtcctgt agaaacccca
     7381 acccgtgaaa tcaaaaaact cgacggcctg tgggcattca gtctggatcg cgaaaactgt
     7441 ggaattgatc agcgttggtg ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca
     7501 ggcagtttta acgatcagtt cgccgatgca gatattcgta attatgcggg caacgtctgg
     7561 tatcagcgcg aagtctttat accgaaaggt aagtcttact ctctcttttt tggtctgtat
     7621 ttttaatttt ttgaagtata ctatttgtac tgacgctaat aatctttttt caggttgggc
     7681 aggccagcgt atcgtgctgc gtttcgatgc ggtcactcat tacggcaaag tgtgggtcaa
     7741 taatcaggaa gtgatggagc atcagggcgg ctatacgcca tttgaagccg atgtcacgcc
     7801 gtatgttatt gccgggaaaa gtgtacgtat caccgtttgt gtgaacaacg aactgaactg
     7861 gcagactatc ccgccgggaa tggtgattac cgacgaaaac ggcaagaaaa agcagtctta
     7921 cttccatgat ttctttaact atgccggaat ccatcgcagc gtaatgctct acaccacgcc
     7981 gaacacctgg gtggacgata tcaccgtggt gacgcatgtc gcgcaagact gtaaccacgc
     8041 gtctgttgac tggcaggtac ttcatgcttc aacgtgtaac ttaagagata ctgtgtgaaa
     8101 ttttatattt ccatacattt gcttgacctt tgctttttgt caattttttt ccccttacag
     8161 gtggtggcca atggtgatgt cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca
     8221 actggacaag gcactagcgg gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt
     8281 gaaggttatc tctatgaact gtgcgtcaca gccaaaagcc agacagagtg tgatatctac
     8341 ccgcttcgcg tcggcatccg gtcagtggca gtgaagggcg aacagttcct gattaaccac
     8401 aaaccgttct actttactgg ctttggtcgt catgaagatg cggacttgcg tggcaaagga
     8461 ttcgataacg tgctgatggt gcacgaccac gcattaatgg actggattgg ggccaactcc
     8521 taccgtacct cgcattaccc ttacgctgaa gagatgctcg actgggcaga tgaacatggc
     8581 atcgtggtga ttgatgaaac tgctgctgtc ggctttaacc tctctttagg cattggtttc
     8641 gaagcgggca acaagccgaa agaactgtac agcgaagagg cagtcaacgg ggaaactcag
     8701 caagcgcact tacaggcgat taaagagctg atagcgcgtg acaaaaacca cccaagcgtg
     8761 gtgatgtgga gtattgccaa cgaaccggat acccgtccgc aaggtgcacg ggaatatttc
     8821 gcgccactgg cggaagcaac gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat
     8881 gtaatgttct gcgacgctca caccgatacc atcagcgatc tctttgatgt gctgtgcctg
     8941 aaccgttatt acggatggta tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg
     9001 gaaaaagaac ttctggcctg gcaggagaaa ctgcatcagc cgattatcat caccgaatac
     9061 ggcgtggata cgttagccgg gctgcactca atgtacaccg acatgtggag tgaagagtat
     9121 cagtgtgcat ggctggatat gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt
     9181 gaacaggtat ggaatttcgc cgattttgcg acctcgcaag gcatattgcg cgttggcggt
     9241 aacaagaaag ggatcttcac tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa
     9301 aaacgctgga ctggcatgaa cttcggtgaa aaaccgcagc agggaggcaa acaatgagct
     9361 tgtcaagcag atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
     9421 ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
     9481 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
     9541 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     9601 gtgtcatcta tgttactaga tcgacgctac tagaggatgc acatgtgacc gagggacacg
     9661 aagtgatccg tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
     9721 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
     9781 tccatttgta tgtgccagcc gcctttgcga cgctcaccgg gctggttgcc ctc
//
