LOCUS       sequence-370239-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zea Mays Nuclear Auxin Signaling CoRepressor REL2N91 and pDEST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  AUTHORS   Ramos Baez R, Buckley Y, Yu H, Chen Z, Gallavotti A, Nemhauser JL, 
            Moss BL
  TITLE     A Synthetic Approach Allows Rapid Characterization of the Maize 
            Nuclear Auxin Response Circuit.
  JOURNAL   Plant Physiol. 2020 Apr;182(4):1713-1722. doi: 10.1104/pp.19.01475. 
            Epub 2020 Mar 2.
  PUBMED    32123041
REFERENCE   2  (bases 1 to 6791)
  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"; doi: 
            "10.1104/pp.19.01475"; journalName: "Plant Physiol"; date: 
            "2020-04"; volume: "182"; issue: "4"; pages: "1713-1722"
COMMENT     Sequence Label: pGP4G-REL2N91-ccdB
FEATURES             Location/Qualifiers
     source          1..6791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        296..443
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             444..1118
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        1171..1814
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1708..1731
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1820..1836
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1820..1836
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2146..2270
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2295..2300,2301..2318,2319..2325)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2379..3059
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(2445..2464)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3379..3684
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3563..3582
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3725..3849)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3856..4102
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3872..3890)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4570..4586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4570..4586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4583..4605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4594..4610
                     /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(4618..4624,4625..4642,4643..4648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4663..4684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4801..4818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4972..5560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5052..5071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5731..6522,6523..6591))
                     /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     6354..6373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6592..6696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6764..6782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accagtttaa accatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac
      241 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
      301 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
      361 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
      421 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
      481 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
      541 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
      601 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
      661 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
      721 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
      781 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
      841 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
      901 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
      961 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     1021 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     1081 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     1141 ttaagtattg tttgtgcact tgccagttta tcattatcaa tactcgccat ttcaaagaat
     1201 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
     1261 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
     1321 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
     1381 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
     1441 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
     1501 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
     1561 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
     1621 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
     1681 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
     1741 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
     1801 gaacttagtt tcgacggatt ctagaactag tggatccccc gggacaaaat gtcgtctctt
     1861 agcagggagc tggttttcct catcctgcag ttcctcgacg aggagaagtt caaagagacg
     1921 gtccacaagt tggagcagga gtcgggcttc tacttcaaca tgaagcactt cgaggacctc
     1981 gtgcagggcg gcgagtggga cgaggtggag aagtacctca gcggcttcac caaggtggag
     2041 gacaaccgct actccatgaa gatcttcttt gagatccgca agcaaaagta cctcgaggcc
     2101 cttgataggc atgacagagc cggtggacca ggtggtggac atcaaacaag tttgtacaaa
     2161 aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat
     2221 aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg gcggccgcat
     2281 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt
     2341 aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2401 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2461 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     2521 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     2581 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     2641 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     2701 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2761 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     2821 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     2881 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     2941 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     3001 tacaacagta ctgcgatgag tggcaggcgg ggcgtaatct agaggatccg gcttactaaa
     3061 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     3121 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     3181 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     3241 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     3301 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     3361 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     3421 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     3481 cccctggcca gtgcacgtct gctgtcagat aaagtccccc gtgaacttta cccggtggtg
     3541 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     3601 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     3661 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     3721 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     3781 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     3841 caaagtggtt cgatgtcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     3901 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     3961 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     4021 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     4081 gacgctcgaa ggctttaatt tgtgcaggcc ttttgaaaag caagcataaa agatctaaac
     4141 ataaaatctg taaaataaca agatgtaaag ataatgctaa atcatttggc tttttgattg
     4201 attgtacagg aaaatataca tcgcaggggg ttgactttta ccatttcacc gcaatggaat
     4261 caaacttgtt gaagagaatg ttcacaggcg catacgctac aatgacccga ttcttgctag
     4321 ccttttctcg gtcttgcaaa caaccgccgg cagcttagta tataaataca catgtacata
     4381 cctctctccg tatcctcgta atcattttct tgtatttatc gtcttttcgc tgtaaaaact
     4441 ttatcacact tatctcaaat acacttatta accgctttta ctattatctt ctacgctgac
     4501 agtaatatca aacagtgaca catattaaac acagtggttt ctttgcataa acaccatgtt
     4561 taaaccatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4621 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4801 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5101 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5161 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5401 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5521 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6601 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6661 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6721 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6781 gccctttcgt c
//
