LOCUS       sequence-77473-e        6243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Confers KAN resistance in planta .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  TITLE     Level 1 KAN and BASTA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6243)
  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            "Unpublished"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: pICSL11024 (pICH47732::NOSp-NPTII-OCST)
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        108..287
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    318..372
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             374..1168
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(428..447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1038..1057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1195..1902
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    1946..1970
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2126..3274)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3690..3709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4369..5160,5161..5229))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4992..5011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5230..5334)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5421..6052
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6144..6168
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gagcggagaa ttaagggagt cacgttatga
       61 cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa ccgcaacgtt
      121 gaaggagcca ctgagccgcg ggtttctgga gtttaatgag ctaagcacat acgtcagaaa
      181 ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat atttcttgtc
      241 aaaaatgctc cactgacgtt ccataaattc ccctcggtat ccaattagag tctcatattc
      301 actctcctat ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
      361 atttacaatt accatggttg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      421 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      481 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      541 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      601 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      661 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      721 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      781 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      841 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      901 gcgcatgccc gacggcgagg atctcgtcgt gactcatggc gatgcctgct tgccgaatat
      961 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1021 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1081 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1141 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgctag agtcctgctt
     1201 taatgagata tgcgagacgc ctatgatcgc atgatatttg ctttcaattc tgttgtgcac
     1261 gttgtaaaaa acctgagcat gtgtagctca gatccttacc gccggtttcg gttcattcta
     1321 atgaatatat cacccgttac tatcgtattt ttatgaataa tattctccgt tcaatttact
     1381 gattgtaccc tactacttat atgtacaata ttaaaatgaa aacaatatat tgtgctgaat
     1441 aggtttatag cgacatctat gatagagcgc cacaataaca aacaattgcg ttttattatt
     1501 acaaatccaa ttttaaaaaa agcggcagaa ccggtcaaac ctaaaagact gattacataa
     1561 atcttattca aatttcaaaa gtgccccagg ggctagtatc tacgacacac cgagcggcga
     1621 actaataacg ctcactgaag ggaactccgg ttccccgccg gcgcgcatgg gtgagattcc
     1681 ttgaagttga gtattggccg tccgctctac cgaaagttac gggcaccatt caacccggtc
     1741 cagcacggcg gccgggtaac cgacttgctg ccccgagaat tatgcagcat ttttttggtg
     1801 tatgtgggcc ccaaatgaag tgcaggtcaa accttgacag tgacgacaaa tcgttgggcg
     1861 ggtccagggc gaattttgcg acaacatgtc gaggctcagc cgctgcaatt gtcttctgca
     1921 cgaagtggtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     1981 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     2041 catttgtatg tgcatgccaa ccacagggtt ccccagatca ggcgctggct gctgaacccc
     2101 cagccggaac tgaccccaca aggccctagc gtttgcaatg caccaggtca tcattgaccc
     2161 aggcgtgttc caccaggccg ctgcctcgca actcttcgca ggcttcgccg acctgctcgc
     2221 gccacttctt cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt tccagcttga
     2281 gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca
     2341 gcttgcggta cttctcccat atgaatttcg tgtagtggtc gccagcaaac agcacgacga
     2401 tttcctcgtc gatcaggacc tggcaacggg acgttttctt gccacggtcc aggacgcgga
     2461 agcggtgcag cagcgacacc gattccaggt gcccaacgcg gtcggacgtg aagcccatcg
     2521 ccgtcgcctg taggcgcgac aggcattcct cggccttcgt gtaataccgg ccattgatcg
     2581 accagcccag gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg ccgatagggg
     2641 tgcgcttcgc gtactccaac acctgctgcc acaccagttc gtcatcgtcg gcccgcagct
     2701 cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca
     2761 gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt
     2821 ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa agctgcattt
     2881 ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg cttggcctcg ctgacctgtt
     2941 ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt catagttcct cgcgtgtcga
     3001 tggtcatcga cttcgccaaa cctgccgcct cctgttcaag acgacgcgaa cgctccacgg
     3061 cggccgatgg cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg
     3121 ccgtagcttg ctggaccatc gagccgacgg actggaaggt ttcgcggggc gcacgcatga
     3181 cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt
     3241 gcctgtatgc cttccggtca aacgtccgat tcattcaccc tccttgcggg attgccccga
     3301 ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc gcctggtgcc ttggtgtcca
     3361 gataatccac cttatcggca atgaagtcgg tcccgtagac cgtctggccg tccttctcgt
     3421 acttggtatt ccgaatcttg ccctgcacga ataccagcga ccccttgccc aaatacttgc
     3481 cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa gaagtcggtg cgctcctgct
     3541 tgtcgccggc atcgttgcgc cacatctagg atctgccagg aaccgtaaaa aggccgcgtt
     3601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4021 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4201 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4261 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4321 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4381 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4441 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4501 gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4561 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4621 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4681 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4741 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4801 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4861 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4921 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4981 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5041 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5101 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5161 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5221 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5281 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacgaa
     5341 ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
     5401 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
     5461 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
     5521 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
     5581 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
     5641 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
     5701 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
     5761 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
     5821 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
     5881 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
     5941 acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg
     6001 gccatagacg gccgccagcc cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga
     6061 gatcctgatc tgactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
     6121 tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt
     6181 agacaactta ataacacatt gcggacgttt ttaatgtact ggggtggatg cagtgggccc
     6241 cac
//
