LOCUS       sequence-248255-        5551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector for transcient expression of proteins 
            with C-terminal 3_Avi in plant protoplast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  AUTHORS   Tu X, Mejia-Guerra MK, Valdes Franco JA, Tzeng D, Chu PY, Shen W, 
            Wei Y, Dai X, Li P, Buckler ES, Zhong S
  TITLE     Reconstructing the maize leaf regulatory network using ChIP-seq data
            of 104 transcription factors.
  JOURNAL   Nat Commun. 2020 Oct 9;11(1):5089. doi: 10.1038/s41467-020-18832-8.
  PUBMED    33037196
REFERENCE   2  (bases 1 to 5551)
  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: "Nat 
            Commun."; date: "2020-10-9"; volume: "11(1):5089. doi"; pages: " 
            10.1038/s41467-020-18832-8"
COMMENT     Sequence Label: p35S-GW-3Avi
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..46
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             567..584
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    591..715
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(750..755,756..773,774..780)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             834..1493
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(900..919)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1835..2140
                     /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     2019..2038
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2181..2305)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2328..2372
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2385..2429
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2442..2486
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     terminator      2745..2997
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3064..3081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3235..3823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3315..3334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3994..4785,4786..4854))
                     /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     4617..4636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4855..4959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5027..5045
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5083..5105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5205..5224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5543..5551,1..12)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggtatt aaaatcttaa
       61 taggttttga taaaagcgaa cgtggggaaa cccgaaccaa accttcttct aaactctctc
      121 tcatctctct taaagcaaac ttctctcttg tctttcttgc gtgagcgatc ttcaacgttg
      181 tcagatcgtg cttcggcacc agtacaacgt tttctttcac tgaagcgaaa tcaaagatct
      241 ctttgtggac acgtagtgcg gcgccattaa ataacgtgta cttgtcctat tcttgtcggt
      301 gtggtcttgg gaaaagaaag cttgctggag gctgctgttc agccccatac attacttgtt
      361 acgattctgc tgactttcgg cgggtgcaat atctctactt ctgcttgacg aggtattgtt
      421 gcctgtactt ctttcttctt cttcttgctg attggttcta taagaaatct agtattttct
      481 ttgaaacaga gttttcccgt ggttttcgaa cttggagaaa gattgttaag cttctgtata
      541 ttctgcccaa attcgcgacc ggtatgcatc accatcacca tcatcccatc acaagtttgt
      601 acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      661 tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc
      721 cctcgagggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
      781 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
      841 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
      901 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
      961 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1021 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     1081 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     1141 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     1201 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     1261 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     1321 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     1381 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1441 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
     1501 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     1561 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     1621 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     1681 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     1741 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     1801 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     1861 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     1921 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     1981 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     2041 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     2101 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     2161 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     2221 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     2281 gttcagcttt cttgtacaaa gtggttgggt accctcgagg cggccgcggc ttgaacgaca
     2341 ttttcgaggc ccagaaaatc gagtggcacg agggtgcggg cgctgggctc aatgacatct
     2401 ttgaggcgca aaaaatagaa tggcatgagg gggctggcgc gggccttaac gatatttttg
     2461 aagcacaaaa aatcgaatgg catgaataga tatgaagatg aagatgttaa ctctggtttc
     2521 attaaatttt ctttagtttg aatttactgt tattcggtgt gcatttctat gtttggtgag
     2581 cggttttctg tgctcagagt gtgtttattt tatgtaattt aatttctttg tgagctcctg
     2641 tttagcaggt cgtcccttca gcaaggacac aaaaagattt taattttatt aaaaaaaaaa
     2701 aaaaaaaaag accgggaatt cgatatcaag cttatcgacc tgcagatcgt tcaaacattt
     2761 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     2821 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     2881 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     2941 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatctct
     3001 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3061 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3661 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4921 tgaatgtatt tagaaaaata aacaaatggg ggttccgcgc acatttcccc gaaaagtgcc
     4981 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5041 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5101 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5161 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5221 tgtactgaga gtgcaccata tgcggccagt gaattgttaa ttaagaattc gagctccacc
     5281 gcggaaacct cctcggattc cattgcccag ctatctgtca ctttattgag aagatagtgg
     5341 aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag
     5401 atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     5461 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg
     5521 taagggatga cgcacaatcc cactatcctt c
//
