LOCUS       sequence-379789-        4784 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 4784)
  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 4784)
  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-35S
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..529
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     472..492
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        725..827
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             858..1163
                     /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     1042..1061
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     terminator      1251..1503
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1521..1537)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1521..1537)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1574..1594)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1574..1592)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1613..1629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1613..1629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1626..1648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1637..1653
                     /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(1661..1667,1668..1685,1686..1691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1706..1727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1844..1861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1966..1990
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2081..2669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2161..2180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2840..3655)
                     /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(2840..3655)
                     /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     3563..3582
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3647..3666
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3946..4381
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4508..4530
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     primer_bind     4680..4702
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4694..4711
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4695..4711
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4721..4740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4721..4739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4764..4780
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4765..4781
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catggagtca aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca
       61 gttcatacag agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga
      121 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
      181 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
      241 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca
      301 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg
      361 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
      421 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
      481 gcaagaccct tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct
      541 tgacagtggt ctctgtccag tcctggcctc gtcggccatt ctcgactaag ttggcagcat
      601 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa
      661 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag
      721 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
      781 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaactt ttgctgacga
      841 gaacagggac tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
      901 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc tgggcgacgg atggtgatcc
      961 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
     1021 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtatccg
     1081 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     1141 acctgatgtt ctggggaata taaatgtcag gctcccttat acacagggcc agtctggcca
     1201 cttgtggtct gctgagaccg tgtgaattgg tgaccagctc gaatttcccc gatcgttcaa
     1261 acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca
     1321 tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat
     1381 ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa
     1441 acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag
     1501 atcgggcctg cagcccgggg gatccactag ttctagagcg gccgccaccg cggtggagct
     1561 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     1621 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     1681 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1741 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1801 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1861 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1921 tatccacaga atcaggggat aacgcaggaa agaacatgaa ggccttgaca ggatatattg
     1981 gcgggtaaac taagtcgctg tatgtgtttg tttgagatct catgtgagca aaaggccagc
     2041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2221 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaga agagttggta
     2581 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2761 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2821 tgtaacattg gtctagtgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2881 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2941 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     3001 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     3061 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     3121 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     3181 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     3241 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     3301 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag
     3361 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     3421 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     3481 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cggtagattg
     3541 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     3601 tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcataacac
     3661 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     3721 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     3781 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg
     3841 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg
     3901 gctccctcac tttctggctg gatgatgggg cgattcaggc gatccccatc caacagcccg
     3961 ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt atccacgtga
     4021 aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct ccaaaaacta
     4081 tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg tgaaatcgct
     4141 aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat agccctttca
     4201 gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt agtatgttca
     4261 attagctttt caattatgaa tatatatatc aattattggt cgcccttggc ttgtggacaa
     4321 tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca ccgtcgagcg
     4381 cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt ttttttttga
     4441 aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga tccccggaat
     4501 tagatcttgg caggatatat tgtggtgtaa cgttaacatt aacgcttaca atttccattc
     4561 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     4621 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     4681 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     4741 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gata
//
