LOCUS       sequence-223588-       10264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non binary level 2 vector adapted for MoClo modular cloning useful 
            for particle bombardment transformation of plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10264)
  TITLE     Non binary MoClo adapted vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10264)
  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            "Unpublished"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: MoClo adapted pUC57-Kan Level 2
FEATURES             Location/Qualifiers
     source          1..10264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      6093..6102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    8096..8117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8234..8251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8405..8993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8485..8504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9171..9980)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9894..9913
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9972..9991
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(10122..10144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggtaccgcac ggtctgccaa tccgacggag gtttatgaat tttccacctt ttccacaagc
       61 tcaactagta ttaacgatgt ggatttagca aaaaaaacct gtaaccctaa atgtaaaata
      121 acgggtaagc ctgccaacca tgttatggca gattaagcgt ctttttgaag ggcaccgcat
      181 ctttcgcgtt gccgtaaatg tatccgttta gcaataagga aattcttaat gaataatccg
      241 tcgttactca atcatgcggt cgaaacgatg gcagttggct cgaaaagttt tgcgacagcc
      301 tcaaagttat ttgatgcaaa aacccggcgc agcgtactga tgctctacgc ctggtgccgc
      361 cattgtgacg atgttattga cgatcagacg ctgggctttc aggcccggca gcctgcctta
      421 caaacgcccg aacaacgtct gatgcaactt gagatgaaaa cgcgccaggc ctatgcagga
      481 tcgcagatgc acgaaccggc gtttgcggct tttcgggaag tggctatggc tcatgatatc
      541 gccccggctt acgcgtttga tcatctggaa ggcttcgcca tggatgtacg cgaagcgcaa
      601 tacagccaac tggatgatac gctgcgctat tgctatcacg ttgcaggcgt tgtcggcttg
      661 atgatggcgc aaatcatggg cgtgcgggat aacgccacgc tggaccgcgc ctgtgacctt
      721 gggctggcat ttcagttgac caatattgct cgcgatattg tggacgatgc gcatgcgggc
      781 cgctgttatc tgccggcaag ctggctggag catgaaggtc tgaacaaaga gaattatgcg
      841 gcacctgaaa accgtcaggc gctgagccgt atcgcccgtc gtttggtgca ggaagcagaa
      901 ccttactatt tgtctgccac agccggcctg gcagggttgc ccctgcgttc cgcctgggca
      961 atcgctacgg cgaagcaggt ttaccggaaa ataggtgtca aagttgaaca ggccggtcag
     1021 caagcctggg atcagcggca gtcaacgacc acgcccgaaa aattaacgct gctgctggcc
     1081 gcctctggtc aggcccttac ttcccggatg cgggctcatc ctccccgccc tgcgcatctc
     1141 tggcagcgcc cgctctaggc ttactataag gatgtttgca atgaaaccaa ctacggtaat
     1201 tggtgcaggc ttcggtggcc tggcactggc aattcgtcta caagctgcgg ggatccccgt
     1261 cttactgctt gaacaacgtg ataaacccgg cggtcgggct tatgtctacg aggatcaggg
     1321 gtttaccttt gatgcaggcc cgacggttat caccgatccc agtgccattg aagaactgtt
     1381 tgcactggca ggaaaacagt taaaagagta tgtcgaactg ctgccggtta cgccgtttta
     1441 ccgcctgtgt tgggagtcag ggaaggtctt taattacgat aacgatcaaa cccggctcga
     1501 agcgcagatt cagcagttta atccccgcga tgtcgaaggt tatcgtcagt ttctggacta
     1561 ttcacgcgcg gtgtttaaag aaggctatct aaagctcggt actgtccctt ttttatcgtt
     1621 cagagacatg cttcgcgccg cacctcaact ggcgaaactg caggcatgga gaagcgttta
     1681 cagtaaggtt gccagttaca tcgaagatga acatctgcgc caggcgtttt ctttccactc
     1741 gctgttggtg ggcggcaatc ccttcgccac ctcatccatt tatacgttga tacacgcgct
     1801 ggagcgtgag tggggcgtct ggtttccgcg tggcggcacc ggcgcattag ttcaggggat
     1861 gataaagctg tttcaggatc tgggtggcga agtcgtgtta aacgccagag tcagccatat
     1921 ggaaacgaca ggaaacaaga ttgaagccgt gcatttagag gacggtcgca ggttcctgac
     1981 gcaagccgtc gcgtcaaatg cagatgtggt tcatacctat cgcgacctgt taagccagca
     2041 ccctgccgcg gttaagcagt ccaacaaact gcagactaag cgcatgagta actctctgtt
     2101 tgtgctctat tttggtttga atcaccatca tgatcagctc gcgcatcaca cggtttgttt
     2161 cggcccgcgt taccgcgagc tgattgacga aatttttaat catgatggcc tcgcagagga
     2221 cttctcactt tatctgcacg cgccctgtgt cacggattcg tcactggcgc ctgaaggttg
     2281 cggcagttac tatgtgttgg cgccggtgcc gcatttaggc accgcgaacc tcgactggac
     2341 ggttgagggg ccaaaactac gcgaccgtat ttttgcgtac cttgagcagc attacatgcc
     2401 tggcttacgg agtcagctgg tcacgcaccg gatgtttacg ccgtttgatt ttcgcgacca
     2461 gcttaatgcc tatcatggct cagccttttc tgtggagccc gttcttaccc agagcgcctg
     2521 gtttcggccg cataaccgcg ataaaaccat tactaatctc tacctggtcg gcgcaggcac
     2581 gcatcccggc gcaggcattc ctggcgtcat cggctcggca aaagcgacag caggtttgat
     2641 gctggaggat ctgatatgag cttttaccca ggagacagag aatgcaaccg cattatgatc
     2701 tgattctcgt gggggctgga ctcgcgaatg gccttatcgc cctgcgactt cagcagcagc
     2761 aacctgatat gcgtattttg cttatcgacg ccgcacccca ggcgggcggg aatcatacgt
     2821 ggtcatttca ccacgatgat ttgactgaga gccaacatcg ttggatagct ccgctggtgg
     2881 ttcatcactg gcccgactat caggtacgct ttcccacacg ccgtcgtaag ctgaacagcg
     2941 gctacttttg tattacttct cagcgtttcg ctgaggtttt acagcgacag tttggcccgc
     3001 acttgtggat ggataccgcg gtcgcagagg ttaatgcgga atctgttcgg ttgaaaaagg
     3061 gtcaggttat cggtgcccgc gcggtgattg acgggcgggg ttatgcggca aattcagcac
     3121 tgagcgtggg cttccaggcg tttattggcc aggaatggcg attgagtcac ccgcatggtt
     3181 tatcatctcc gattatcatg gatgccacgg tcgatcagca aaatggttat cgcttcgtgt
     3241 acagcctgcc gctctcgccg accagattgt taattgaaga tacgcactat attgataatg
     3301 cgacattaga tcctgaatgc gcgcggcaaa atatttgcga ctatgccgcg caacagggtt
     3361 ggcagcttca gacactgctg cgagaagaac agggcgcctt acccattact ctgtcgggca
     3421 atgccgacgc attctggcag cagcgccccc tggcctgtag tggattacgt gccggtctgt
     3481 tccatcctac caccggctat tcactgccgc tggcggttgc cgtggccgac cgcctgagtg
     3541 cacttgatgt ctttacgtcg gcctcaattc accatgccat tacgcatttt gcccgcgagc
     3601 gctggcagca gcagggcttt ttccgcatgc tgaatcgcat gctgttttta gccggacccg
     3661 ccgattcacg ctggcgggtt atgcagcgtt tttatggttt acctgaagat ttaattgccc
     3721 gtttttatgc gggaaaactc acgctgaccg atcggctacg tattctgagc ggcaagccgc
     3781 ctgttccggt attagcagca ttgcaagcca ttatgacgac tcatcgttaa gcttcagagg
     3841 aggaattaag taatgagttt tgatattgcc aaatacccga ccctggcact ggtcgactcc
     3901 acccaggagt tacgactgtt gccgaaagag agtttaccga aactctgcga cgaactgcgc
     3961 cgctatttac tcgacagcgt gagccgttcc agcgggcact tcgcctccgg gctgggcacg
     4021 gtcgaactga ccgtggcgct gcactatgtc tacaacaccc cgtttgacca attgatttgg
     4081 gatgtggggc atcaggctta tccgcataaa attttgaccg gacgccgcga caaaatcggc
     4141 accatccgtc agaaaggcgg tctgcacccg ttcccgtggc gcggcgaaag cgaatatgac
     4201 gtattaagcg tcgggcattc atcaacctcc attagtgccg gaattggtat tgcggttgct
     4261 gccgaaaaag aaggcaaaaa tcgccgcacc gtctgtgtca ttggcgatgg cgcgattacc
     4321 gcaggcatgg cgtttgaagc gatgaatcac gcgggcgata tccgtcctga tatgctggtg
     4381 attctcaacg acaatgaaat gtcgatttcc gaaaatgtcg gcgcgctcaa caaccatctg
     4441 gcacagctgc tttccggtaa gctttactct tcactgcgcg aaggcgggaa aaaagttttc
     4501 tctggcgtgc cgccaattaa agagctgctc aaacgcaccg aagaacatat taaaggcatg
     4561 gtagtgcctg gcacgttgtt tgaggagctg ggctttaact acatcggccc ggtggacggt
     4621 catgatgtgc tggggcttat caccacgcta aagaacatgc gcgacctgaa aggcccgcag
     4681 ttcctgcata tcatgaccaa aaaaggtcgt ggttatgaac cggcagaaaa agacccgatc
     4741 actttccatg ccgtgctaaa tttgatccct ccagcggttg tttgccgaaa agtagcggcg
     4801 gtttgccgag ctattcaaaa atctttggcg actggttgtg cgaaacggca gcgaaagaca
     4861 acaagctgat ggcgattact ccggcgatgc gtgaaggttc cggcatggtc gagttttcac
     4921 gtaaattccc ggatcgctac ttcgacgtgg caattgccga gcaacatgcg gtgacctttg
     4981 ctgcgggtct ggcgattggt gggtacaaac ccattgtcgc gatttactcc actttcctgc
     5041 aacgcgccta tgatcaggtg ctgcatgacg tggcgattca aaagcttccg gtcctgttcg
     5101 ccatcgaccg cgcgggcatt gttggtgctg acggtcaaac ccatcagggt gcttttgatc
     5161 tctcttacct gcgctgcata ccggaaatgg tcattatgac cccgagcgat gaaaacgaat
     5221 gtcgccagat gctctatacc ggctatcact ataacgatgg cccgtcagcg gtgcgctacc
     5281 cgcgtggcaa cgcggtcggc gtggaactga cgccgctgga aaaactacca attggcaaag
     5341 gcattgtgaa gcgtcgtggc gagaaactgg cgatccttaa ctttggtacg ctgatgccag
     5401 aagcggcgaa agtcgccgaa tcgctgaacg ccacgctggt cgatatgcgt tttgtgaaac
     5461 cgcttgatga agcgttaatt ctggaaatgg ccgccagcca tgaagcgctg gtcaccgtag
     5521 aagaaaacgc cattatgggc ggcgcaggca gcggcgtgaa cgaagtgctg atggcccatc
     5581 gtaaaccagt acccgtgctg aacattggcc tgccggactt ctttattccg caaggaactc
     5641 aggaagaaat gcgcgccgaa ctcggcctcg atgccgctgg tatggaagcc aaaatcaagg
     5701 cctggctggc ataagctttg tgtgaggaga tgtcgaatga gcgcacatgc cctgcccaag
     5761 gcagatctga ccgccaccag cctgatcgta tcgggcggca tcatcgccgc ttggctggcc
     5821 ctgcatgtgc atgcgctgtg gtttctggac gcagcggcgc atcccatcct ggcgatcgca
     5881 aatttcctgg ggctgacctg gctgtcggtc ggattgttca tcatcgcgca tgacgcgatg
     5941 cacgggtcgg tggtgccggg gcgtccgcgc gccaatgcgg cgatgggcca gcttgtcctg
     6001 tggctgtatg ccggattttc gtggcgcaag atgatcgtca agcacatggc ccatcaccgc
     6061 catgccggaa ccgacgacga ccccgatttc gaccatggcg gcccggtccg ctggtacgcc
     6121 cgcttcatcg gcacctattt cggctggcgc gaggggctgc ttctgcccgt catcgtgacg
     6181 gtctatgcgc tgatccttgg ggatcgctgg atgtacgtgg tattctggcc gctgccgtcg
     6241 atcctggcgt cgatccagct gttcgtgttc ggcacctggc tgccgcaccg ccccggccac
     6301 gacgcgttcc cggaccgcca caatgcgcgg tcgtcgcgga tcagcgaccc cgtgtcgctg
     6361 ctgacctgtt ttcactttgg cggttatcat cacgaacacc acctgcaccc gacggtgccg
     6421 tggtggcgcc tgcccagcac ccgcaccaag ggcgacaccg cataagcttg agctgaggag
     6481 attttgaatg caaacggaac acgtcatttt attgaatgca cagggagttc ccacgggtac
     6541 gctggaaaag tatgccgcac acacggcaga cacccgctta catctcgcgt tctccagttg
     6601 gctgtttaat gccaaaggac aattattagt tacccgccgc gcactgagca aaaaagcatg
     6661 gcctggcgtg tggactaact cggtttgtgg gcacccacaa ctgggagaaa gcaacgagga
     6721 cgcagtgatc cgccgttgcc gttatgagct tggcgtggaa attacgcctc ctgaatctat
     6781 ctatcctgac tttcgctacc gcgccaccga tccgagtggc attgtggaaa atgaagtgtg
     6841 tccggtattt gccgcacgca ccactagtgc gttacagatc aatgatgatg aagtgatgga
     6901 ttatcaatgg tgtgatttag cagatgtatt acacggtatt gatgcaacgc cgtgggcgtt
     6961 cagtccgtgg atggtgatgc aggcgacaaa tcgcgaagcc agaaaacgat tatctgcatt
     7021 tacccagctt aaataagctt tgcctcagga tttttgaaat gacggtctgc gcaaaaaaac
     7081 acgttcatct cactcgcgat gctgcggagc agttactggc tgatattgat cgacgccttg
     7141 atcagttatt gcccgtggag ggagaacggg atgttgtggg tgccgcgatg cgtgaaggtg
     7201 cgctggcacc gggaaaacgt attcgcccca tgttgctgtt gctgaccgcc cgcgatctgg
     7261 gttgcgctgt cagccatgac ggattactgg atttggcctg tgcggtggaa atggtccacg
     7321 cggcttcgct gatccttgac gatatgccct gcatggacga tgcgaagctg cggcgcggac
     7381 gccctaccat tcattctcat tacggagagc atgtggcaat actggcggcg gttgccttgc
     7441 tgagtaaagc ctttggcgta attgccgatg cagatggcct cacgccgctg gcaaaaaatc
     7501 gggcggtttc tgaactgtca aacgccatcg gcatgcaagg attggttcag ggtcagttca
     7561 aggatctgtc tgaaggggat aagccgcgca gcgctgaagc tattttgatg acgaatcact
     7621 ttaaaaccag cacgctgttt tgtgcctcga tgcagatggc ctcgattgtt gcgaatgcct
     7681 ccagcgaagc gcgtgattgc ctgcatcgtt tttcacttga tcttggtcag gcatttcaac
     7741 tgctggacga tttgaccgat ggcatgaccg acaccggtaa ggatagcaat caggacgccg
     7801 gtaaatcgac gctggtcaat ctgttaggcc cgagggcggt tgaagaacgt ctgagacaac
     7861 atcttcagct tgccagtgag catctctctg cggcctgcca acacgggcac gccactcaac
     7921 attttattca ggcctggttt gacaaaaaac tcgctgccgt cagttaagct tgcaagaatt
     7981 caagcttgga gtacgttaat tagttgtctg ttttaagccc cgtgctctcc gccggggctt
     8041 ttttattaaa aaccactcaa gtggaagaca agggacacga agtgtggggt gcctaatgag
     8101 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8161 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8221 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8281 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8341 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8401 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8461 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8521 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8581 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8641 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8701 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8761 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8821 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8881 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8941 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9001 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9061 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9121 ttaaatcaag cccaatctga ataatgttac aaccaattaa ccaattctga ttagaaaaac
     9181 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     9241 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     9301 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     9361 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     9421 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     9481 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     9541 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     9601 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     9661 acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta
     9721 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     9781 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     9841 gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga
     9901 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt
     9961 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
    10021 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
    10081 ggccagagct gcatcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
    10141 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
    10201 gcgtcagcgg gtgttggcgg gtgtcggggc tggccactct gtgccttgtc ttccagtggt
    10261 atgg
//
