LOCUS       sequence-68094-e        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary vector for Arabidopsis transformation with BsaI Golden Gate 
            sites and Kanamycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Emami S, Yee MC, Dinneny JR
  TITLE     A robust family of Golden Gate Agrobacterium vectors for plant 
            synthetic biology.
  JOURNAL   Front Plant Sci. 2013 Sep 2;4:339. doi: 10.3389/fpls.2013.00339. 
            eCollection 2013.
  PUBMED    24032037
REFERENCE   2  (bases 1 to 6013)
  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: "Front Plant
            Sci."; date: "2013-09-2"; volume: "4:339. doi"; pages: " 
            10.3389/fpls.2013.00339. eCollection 2013"
COMMENT     Sequence Label: pGoldenGate-SE9
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(57..309)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(326..1120)
                     /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
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(437..456)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1047..1066
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1141..1324)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(1177..1356)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             complement(1518..1841)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1523..1542
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1736..1758
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1750..1767
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1751..1767
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1768..1824
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1837..1853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1861..1877
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1885..1891,1892..1909,1910..1915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1930..1951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    2187..2363
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(2427..2447)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2427..2445)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2466..2482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2490..2506
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2514..2520,2521..2538,2539..2544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2559..2580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2819..2843
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3014..3033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3613..4401)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5453..5888
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cttggcagga tatattgtgg tgtaacgttg gcgcgccatc agcttgcatg ccggtcgatc
       61 tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt
      121 ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa
      181 taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca gaaattatat
      241 gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt
      301 tgaacgatct gcttgactct accgctcaga agaactcgtc aagaaggcga tagaaggcga
      361 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
      421 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
      481 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
      541 gcaagcaggc atcgccatgg gtcacgacga gatcatcgcc gtcgggcatg cgcgccttga
      601 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
      661 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
      721 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
      781 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
      841 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
      901 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
      961 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     1021 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     1081 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat ccagatccgg
     1141 tgcagattat ttggattgag agtgaatatg agactctaat tggataccga ggggaattta
     1201 tggaacgtca gtggagcatt tttgacaaga aatatttgct agctgatagt gaccttaggc
     1261 gacttttgaa cgcgcaataa tggtttctga cgtatgtgct tagctcatta aactccagaa
     1321 acccgcggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc
     1381 ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg ctcgagagta
     1441 agagacccgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1501 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     1561 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     1621 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     1681 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     1741 tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga
     1801 gtcgacctgc aggcatgcag gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2041 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2101 gctgcggcga gcggtatcgg tctcatggaa agcttgtcgg tacgctgaaa tcaccagtct
     2161 ctctctacaa atctatctct ctctattttc tccataaata atgtgtgagt agtttcccga
     2221 taagggaaat tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt
     2281 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
     2341 aatccagtac taaaatccag atcgatgggc tagagcggac tagttctaga gcggccgcca
     2401 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     2461 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2521 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2581 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2641 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2701 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2761 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gaaggccttg
     2821 acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga
     2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3421 agaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaat
     3601 tcttgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtgaacaaa
     3661 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat aagcctgcct
     3721 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     3781 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     3841 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     3901 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     3961 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     4021 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     4081 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     4141 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     4201 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     4261 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     4321 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     4381 ggcgatcacc gcttccctca tgatgtttaa ctcctgaatt aagccgcgcc gcgaagcggt
     4441 gtggcttgaa tgaattgtta ggcgtcatcc tgtgctcccg agaaccagta ccagtacatc
     4501 gctgtttcgt tcgagacttg aggtctagtt ttatacgtga acaggtcaat gccgccgaga
     4561 gtaaagccac atttgcgtac aaattgcagg caggtacatt gttcgtttgt gtctctaatc
     4621 gtatgccaag gagctgtctg cttagtgccc actttttcgc aaaattcgat gagactgtgc
     4681 gcgactcctt tgcctcggtg cgtgtgcgac acaacaatgt gttcgataga ggctagatcg
     4741 ttcccatgtt gagttgagtt caatcttccc gacaagctct tggtcgatga atgtgccata
     4801 gcaagcagag tcttcatcag agtcatcatc cgagatgtaa tccttccggt aggggctcac
     4861 acttctggta gatagttcaa agccttggtc ggataggtgc acatcgaaca cttcacgaac
     4921 aatgaaatgg ttctcagcat ccaatgtttc cgccacctgc tcagggatca ccgaaatctt
     4981 catatgacgc ctaacgcctg gcacagcgga tcgcaaacct ggcgcggctt ttggcacaaa
     5041 aggcgtgaca ggtttgcgaa tccgttgctg ccacttgttt aatagactgg atggaggcgg
     5101 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5161 aatctggagc cggtgagcgt ggatctcgcg gtatcattgc agcactgcca tgatgatata
     5221 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata
     5281 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     5341 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     5401 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat ccccatccaa
     5461 cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag ggtagttatc
     5521 cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg gcccgctcca
     5581 aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga
     5641 aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc
     5701 cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac agcaagtagt
     5761 atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc ccttggcttg
     5821 tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg
     5881 tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt
     5941 tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc
     6001 ccggaattag act
//
