LOCUS       sequence-247702-        7836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty control for pGAPi-based RNAi silencing in tandem with APT 
            transcript segment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  AUTHORS   Orr RG, Foley SJ, Sherman CA, Abreu I, Galotto G, Liu B, 
            Gonzalez-Guerrero M, Vidali L
  TITLE     Robust survival-based RNAi of gene families using in tandem 
            silencing of adenine phosphoribosyltransferase.
  JOURNAL   Plant Physiol. 2020 Aug 6. pii: pp.20.00865. doi: 
            10.1104/pp.20.00865.
  PUBMED    32764132
REFERENCE   2  (bases 1 to 7836)
  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 
            Physiol. 2020 Aug 6. pii: pp.20.00865. doi: 10.1104/pp.20.00865."
COMMENT     Sequence Label: pAPi
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2236..2488
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2521..2866
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2809..2829
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2896..3921
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2896..3921
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3991..4167
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(4721..4737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4721..4737)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4734..4756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4745..4761
                     /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(4769..4775,4776..4793,4794..4799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4814..4835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4952..4969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5123..5711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5203..5222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5882..6673,6674..6742))
                     /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     6505..6524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6743..6847)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccnccccc tctctacctt ctctagatcg gcgttccggt ccatggttag ggcccggtag
       61 ttctacttct gttcatgttt gtgttagatc cgtgtttgtg ttagatccgt gctgctagcg
      121 ttcgtacacg gatgcgacct gtacgtcaga cacgttctga ttgctaactt gccagtgttt
      181 ctctttgggg aatcctggga tggctctagc cgttccgcag acgggatcga tttcatgatt
      241 ttttttgttt cgttgcatag ggtttggttt gcccttttcc tttatttcaa tatatgccgt
      301 gcacttgttt gtcgggtcat cttttcatgc ttttttttgt cttggttgtg atgatgtggt
      361 ctggttgggc ggtcgttcta gatcggagta gaattaattc tgtttcaaac tacctggtgg
      421 atttattaat tttggatctg tatgtgtgtg ccatacatat tcatagttac gaattgaaga
      481 tgatggatgg aaatatcgat ctaggatagg tatacatgtt gatgcgggtt ttactgatgc
      541 atatacagag atgctttttg ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt
      601 tcattcgttc tagatcggag tagaatactg tttcaaacta cctggtgtat ttattaattt
      661 tggaactgta tgtgtgtgtc atacatcttc atagttacga gtttaagatg gatggaaata
      721 tcgatctagg ataggtatac atgttgatgt gggttttact gatgcatata catgatggca
      781 tatgcagcat ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta
      841 tgttttataa ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg
      901 tggatttttt tagccctgcc ttcatacgct atttatttgc ttggtactgt ttcttttgtc
      961 gatgctcacc ctgttgtttg gtgttacttc tgcgccagtg ccaagttaat tcgagctcaa
     1021 gctttaccgt gcgccacttg acacagactg gaaatattct cgttcacaca gtgctagggc
     1081 ttcagggtgt gagccgtggc gcatcttgtg gacttgccgt ggcgagctac ctccacgctc
     1141 ccgcttccgc aattttgttg cagtgctttg gattctcgtg gttgctcgtc tcaggaagtg
     1201 aagatgtcgg cctccaagga tggcgaccct cggatccagt atatctcgga cagcatccgt
     1261 accattcctg attttcctca caaaggcatt atgttccgag atgtgacgac gttgctgttg
     1321 gatcataagg ctttcaaaga cacgatcgac atctttgttg agcgttaccg ggaccagaag
     1381 gtggacgtca ttgtgggaat tgaagctcga ggtatttaaa ttgattacgt acagatgaac
     1441 atggcatcgt ggtgattgat gaaactgctg ctgtcggctt taacctctct ttaggcattg
     1501 gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga agaggcagtc aacggggaaa
     1561 ctcagcaagc gcacttacag gcgattaaag agctgatagc gcgtgacaaa aaccacccaa
     1621 gcgtggtgat gtggagtatt gccaacgaac cctcgaggga tacccgtccg caaggtgcac
     1681 gggaatattt cgcgccactg gcggaagcaa cgcgtaaact cgacccgacg cgtccgatca
     1741 cctgcgtcaa tgtaatgttc tgcgacgctc acaccgatac catcagcgat ctctttgatg
     1801 cctaggtacg taatcagttt aaatacctcg agcttcaatt cccacaatga cgtccacctt
     1861 ctggtcccgg taacgctcaa caaagatgtc gatcgtgtct ttgaaagcct tatgatccaa
     1921 cagcaacgtc gtcacatctc ggaacataat gcctttgtga ggaaaatcag gaatggtacg
     1981 gatgctgtcc gagatatact ggatccgagg gtcgccatcc ttggaggccg acatcttcac
     2041 ttcctgagac gagcaaccac gagaatccaa agcactgcaa caaaattgcg gaagcgggag
     2101 cgtggaggta gctcgccacg gcaagtccac aagatgcgcc acggctcaca ccctgaagcc
     2161 ctagcactgt gtgaacgaga atatttccag tctgtgtcaa gtggcgcacg gtaaagcttg
     2221 gtacccctac tccaagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     2281 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
     2341 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     2401 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     2461 cgcgcggtgt catctatgtt actagatcgg ggatacagtc tcagaagacc aaagggctat
     2521 tgagactttt caacaaaggg taatttcggg aaacctcctc ggattccatt gcccagctat
     2581 ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
     2641 cgataaagga aaggctatca ttcaagatgc ctctgccgac agtggtccca aagatggacc
     2701 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
     2761 ggattgatgt gacatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
     2821 agacccttcc tctatataag gaagttcatt tcatttggag aggacagccc aagcatggag
     2881 aaccgacctg caggcatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     2941 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     3001 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     3061 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     3121 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     3181 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     3241 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     3301 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     3361 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     3421 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     3481 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     3541 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     3601 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     3661 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     3721 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     3781 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     3841 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     3901 cgtccgaggg caaaggaatg agcgggactc tggggttcgg acggtacgct gaaatcacca
     3961 gtctctctct acaaatctat ctctctctat tttctccata aataatgtgt gagtagtttc
     4021 ccgataaggg aaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac
     4081 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     4141 ccaaaatcca gtactaaaat ccagatctcc taaagtccct atagatcttt gtcgtgaata
     4201 taaaccagac acgagacgac taaacctgga gcccagacgc cgttcgaagc tagaagtacc
     4261 gcttaggcag gaggccgtta gggaaaagat gctaaggcag ggttggttac gttgactccc
     4321 ccgtaggttt ggtttaaata tgatgaagtg gacggaagga aggaggaaga caaggaagga
     4381 taaggttgca ggccctgtgc aaggtaagaa gatggaaatt tgatagaggt acgctactat
     4441 acttatacta tacgctaagg gaatgcttgt atttataccc tataccccct aataacccct
     4501 tatcaattta agaaataatc cgcataagcc cccgcttaaa aattggtatc agagccatga
     4561 ataggtctat gaccaaaact caagaggata aaacctcacc aaaatacgaa agagttctta
     4621 actctaaaga taaaagatct ttcaagatca aaactagttc cctcacaccg gtgacgggga
     4681 tcgcatgcga tatctcgaga tctagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4741 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4801 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgtgctca ctgcccgctt
     4861 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4921 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4981 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5041 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5101 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5161 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5221 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5281 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5341 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5401 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5461 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5521 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5581 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5641 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5701 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5761 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5821 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5881 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5941 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6001 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6061 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6121 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6181 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6241 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6301 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6361 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6421 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6481 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6541 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6601 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6661 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6721 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6781 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6841 ttccgcgcac atttccccga aaagtgccta gagataatga gcattgcatg tctaagttat
     6901 aaaaaattac cacatatttt ttttgtcaca cttgtttgaa gtgcagttta tctatcttta
     6961 tacatatatt taaactttac tctacgaata atataatcta tagtactaca ataatatcag
     7021 tgttttagag aatcatataa atgaacagtt agacatggtc taaaggacaa ttgagtattt
     7081 tgacaacagg actctacagt tttatctttt tagtgtgcat gtgttctcct ttttttttgc
     7141 aaatagcttc acctatataa tacttcatcc attttattag tacatccatt tagggtttag
     7201 ggttaatggt ttttatagac taattttttt agtacatcta ttttattcta ttttagcctc
     7261 taaattaaga aaactaaaac tctattttag tttttttatt taataattta gatataaaat
     7321 agaataaaat aaagtgacta aaaattaaac aaataccctt taagaaatta aaaaaactaa
     7381 ggaaacattt ttcttgtttc gagtagataa tgccagcctg ttaaacgccg tcgacgagtc
     7441 taacggacac caaccagcga accagcagcg tcgcgtcggg ccaagcgaag cagacggcac
     7501 ggcatctctg tcgctgcctc tggacccctc tcgagagttc cgctccaccg ttggacttgc
     7561 tccgctgtcg gcatccagaa attgcgtggc ggagcggcag acgtgagccg gcacggcagg
     7621 cggcctcctc ctcctctcac ggcaccggca gctacggggg attcctttcc caccgctcct
     7681 tcgctttccc ttcctcgccc gccgtaataa atagacaccc cctccacacc ctctttcccc
     7741 aacctcgtgt tgttcggagc gcacacacac acaaccagat ctcccccaaa tccacccgtc
     7801 ggcacctccg cttcaaggta cgccgctcgt cctccc
//
