LOCUS       sequence-292691-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone with nourseothricin selection in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  TITLE     Quantitative comparison of specialised leaf promoters
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8203)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pN_empty
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    323..347
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1678..2307
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2736..3809
                     /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      3875..4069
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4305..4327)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4413..4553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4468..4487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4568..4585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4739..5327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4819..4838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5573..6364)
                     /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"
     misc_feature    6885..6909
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7002..7254)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7277..7849)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(7891..8070)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     protein_bind    complement(join(8182..8203,1..3))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggtgatatcc cgcggccatg ctagagtccg caaaaatcac cagtctctct ctacaaatct
       61 atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg
      121 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
      181 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgacctgc
      241 aggcatgcga cgtcgggccc aagcttagct tgagcttgga tcagattgtc gtttcccgcc
      301 ttcagtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga
      361 gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat
      421 ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtacttta aagtacttta
      481 aagtacttta aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
      541 gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
      601 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
      661 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
      721 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
      781 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
      841 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
      901 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
      961 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     1021 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     1081 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     1141 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     1201 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     1261 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     1321 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     1381 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     1441 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc tcaaccgtgc ggctgcatga
     1501 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     1561 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     1621 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     1681 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     1741 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     1801 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     1861 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     1921 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     1981 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     2041 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     2101 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     2161 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     2221 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     2281 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     2341 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     2401 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     2461 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     2521 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     2581 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     2641 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc
     2701 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
     2761 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     2821 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     2881 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     2941 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     3001 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     3061 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     3121 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     3181 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     3241 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     3301 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
     3361 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
     3421 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
     3481 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
     3541 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
     3601 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
     3661 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
     3721 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
     3781 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
     3841 tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac attgggaacc
     3901 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
     3961 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
     4021 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
     4081 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
     4141 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
     4201 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
     4261 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4321 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4381 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     4441 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4501 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     4561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5161 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5401 tgcatgatat atctcccaat ttgtgtaggg cttattatgc acgcttaaaa ataataaaag
     5461 cagacttgac ctgatagttt ggctgtgagc aattatgtgc ttagtgcatc taacgcttga
     5521 gttaagccgc gccgcgaagc ggcgtcggct tgaacgaatt tctagctaga cattatttgc
     5581 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     5641 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     5701 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     5761 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     5821 cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     5881 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     5941 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     6001 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     6061 gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa
     6121 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     6181 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     6241 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     6301 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     6361 ccatgatgtt taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc
     6421 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     6481 actgtacccc aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     6541 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     6601 gcttacgaac cgaacaggct tatgtccact gggttcgtgc ccgaattgat cacaggcagc
     6661 aacgctctgt catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa
     6721 cccggcagct tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc
     6781 cttacaacgg ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt
     6841 gattttgtgc cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt
     6901 ggtgtaaaca aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac
     6961 tgaattaacg ccgaattgaa ttatcagctt gcatgccggt cgatctagta acatagatga
     7021 caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta
     7081 aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta
     7141 catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag
     7201 accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatctgcttg
     7261 actctagggg tcatcattag gggcagggca tgctcatgta gagcgcctgc tcgccgtccg
     7321 aggcggtgcc gtcgtacagg gcggtgtcca ggccgcagag ggtgaacccc atccgccggt
     7381 acgcgtggat cgccggtgcg ttgacgttgg tgacctccag ccagaggtgc ccggcacccc
     7441 gctcgcgggc gaactccgtc gcgagcccca tcaacgcgcg cccgaccccg tgcccccggt
     7501 gctccggggc gacctcgatg tcctcgacgg tcagccggcg gttccagccg gagtacgaga
     7561 cgaccacgaa gcccgccagg tcgccgtcgt ccccgtacgc gacgaacgtc cgagagtccg
     7621 ggtcgccgtc ctccccgtcg tccgattcgt cgtccgattc gtcgtcgggg aacaccttgg
     7681 tcaggggcgg gtccaccggc acctcccgca gggtgaagcc gtccccggtg gcggtgacgc
     7741 ggaagacggt gtcggtggtg aaggacccat ccagtgcctc gatggcctcg gcgtcccccg
     7801 ggacactggt gcggtaccgg taagccgtgt cgtcaagagt ggtacccatg gtagatcccc
     7861 tcgatcgagt tgagagtgaa tatgagactc taattggata ccgaggggaa tttatggaac
     7921 gtcagtggag catttttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt
     7981 tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc
     8041 ggctcagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc
     8101 cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg attgataatt
     8161 cgagctctcc catatggtct cgttcagctt tcttgtacaa agt
//
