LOCUS       sequence-383129-        3953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nimble cloning-compatible vector for plant ectopic expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3953)
  AUTHORS   Yan P, Tuo D, Shen W, Deng H, Zhou P, Gao X
  TITLE     A Nimble Cloning-compatible vector system for high-throughput gene 
            functional analysis in plants.
  JOURNAL   Plant Commun. 2023 Mar 13;4(2):100471. doi: 
            10.1016/j.xplc.2022.100471. Epub 2022 Nov 9.
  PUBMED    36352791
REFERENCE   2  (bases 1 to 3953)
  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 
            Commun."; date: "2023-03-13"; volume: "4(2):100471. doi"; pages: " 
            10.1016/j.xplc.2022.100471. Epub 2022 Nov 9"
COMMENT     Sequence Label: pNC-Green
FEATURES             Location/Qualifiers
     source          1..3953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        181..283
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             314..619
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     498..517
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(690..706)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(690..706)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(743..763)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(743..761)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(782..798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(782..798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(795..817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    806..822
                     /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(830..836,837..854,855..860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    875..896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1013..1030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1135..1159
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1250..1838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1330..1349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2009..2824)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2009..2824)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2732..2751
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2816..2835
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3115..3550
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    3677..3699
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     primer_bind     3849..3871
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3863..3880
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3864..3880
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3890..3909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3890..3908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3933..3949
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3934..3950
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agtggtctct gtccagtcct ggcctcgtcg gccattctcg actaagttgg cagcatcacc
       61 cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
      121 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt
      181 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
      241 attttttgag ttatcgagat tttcaggagc taaggaagct aaacttttgc tgacgagaac
      301 agggactggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
      361 gtttgtggat gtacagagtg atattattga cacgcctggg cgacggatgg tgatccccct
      421 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
      481 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tatccgttat
      541 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
      601 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agggccagtc tggccacttg
      661 tggtctgctg agacccctgc agcccggggg atccactagt tctagagcgg ccgccaccgc
      721 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
      781 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      841 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      901 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      961 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgaag gccttgacag
     1141 gatatattgg cgggtaaact aarkcgctgt atgtgtttgt ttgagatctc atgtgagcaa
     1201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa
     1741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1981 gtatatatgt gtaacattgg tctagtgatt agaaaaactc atcgagcatc aaatgaaact
     2041 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2101 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     2161 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2221 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     2281 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2341 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     2401 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     2461 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg
     2521 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     2581 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     2641 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     2701 ggtagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     2761 cagcatccat gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc
     2821 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     2881 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     2941 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc
     3001 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca
     3061 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcg atccccatcc
     3121 aacagcccgc cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta
     3181 tccacgtgaa accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc
     3241 caaaaactat ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt
     3301 gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata
     3361 gccctttcag atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta
     3421 gtatgttcaa ttagcttttc aattatgaat atatatatca attattggtc gcccttggct
     3481 tgtggacaat gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac
     3541 cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt
     3601 tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat
     3661 ccccggaatt agatcttggc aggatatatt gtggtgtaac gttaacatta acgcttacaa
     3721 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3781 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3841 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     3901 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ata
//
