LOCUS       sequence-297763-        8281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mPing Yeast Transposition Assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8281)
  AUTHORS   Johnson A, Mcassey E, Diaz S, Reagin J, Redd PS, Parrilla DR, Nguyen
            H, Stec A, McDaniel LAL, Clemente TE, Stupar RM, Parrott WA, Hancock
            CN
  TITLE     Development of mPing-based activation tags for crop insertional 
            mutagenesis.
  JOURNAL   Plant Direct. 2021 Jan 19;5(1):e00300. doi: 10.1002/pld3.300. 
            eCollection 2021 Jan.
  PUBMED    33506165
REFERENCE   2  (bases 1 to 8281)
  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 
            Direct."; date: "2021-01-19"; volume: "5(1):e00300. doi"; pages: " 
            10.1002/pld3.300. eCollection 2021 Jan"
COMMENT     Sequence Label: pWL89A mmPing20
FEATURES             Location/Qualifiers
     source          1..8281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        277..652
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     primer_bind     complement(2976..2993)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2976..2992)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2985..3007)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3749..4911
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      4074..4911
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5246..6049)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6050..6265)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6290..6308)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6376..6480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6481..6549,6550..7341)
                     /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     complement(6699..6718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7512..8100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8001..8020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8254..8271
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgactct agaggatctg aattaattct tgaataatac ataacttttc
      301 ttaaaagaat caaagacaga taaaatttaa gagatattaa atattagtga gaagccgaga
      361 attttgtaac accaacataa cactgacatc tttaacaact tttaattatg atacatttct
      421 tacgtcatga ttgattatta cagctatgct gacaaatgac tcttgttgca tggctacgaa
      481 ccgggtaata ctaagtgatt gactcttgct gaccttttat taagaactaa atggacaata
      541 ttatggagca tttcatgtat aaattggtgc gtaaaatcgt tggatctctc ttctaagtac
      601 atcctactat aacaatcaag aaaaacaaga aaatcggacc tcgagatcaa gtatggattc
      661 tagaacagtt ggtatattag gagggggaca attgggacgt atgattgttg aggcagcaaa
      721 caggctcaac attaagacgg taatactaga tgctgaaaat tctcctgcca aacaaataag
      781 caactccaat gaccacgtta atggctcctt ttccaatcct cttgatatcg aaaaactagc
      841 tgaaaaatgt gatgtgctaa cgattgagat tgagcatgtt gatgttccta cactaaagaa
      901 tcttcaagta aaacatccca aattaaaaat ttacccttct ccagaaacaa tcagattgat
      961 acaagacaaa tatattcaaa aagagcattt aatcaaaaat ggtatagcag ttacccaaag
     1021 tgttcctgtg gaacaagcca gtgagacgtc cctattgaat gttggaagag atttgggttt
     1081 tccattcgtc ttgaagtcga ggactttggc atacgatgga agaggtaact tcgttgtaaa
     1141 gaataaggaa atgattccgg aagctttgga agtactgaag gatcgtcctt tgtacgccga
     1201 aaaatgggca ccatttacta aagaattagc agtcatgatt gtgaggtctg ttaggccagt
     1261 cacaatggct agtgtcattg cacggctacc caaaatatta taccatcttc tttcaaatga
     1321 aatcttttat gaaacaatcc ccacagtgga ggggtttcac tttgacgttt ccatgactat
     1381 gcatagcatt taattggtac aagttgctgg gatcatttgt acccataatc cggcgcggcg
     1441 cgggagaatg cagaggtcgc acggcggagg cggacgcaag agatccggtg aatgaaacga
     1501 atcggcctca acgggggttt cgctctgtta ccgaggactt ggaaacgacg ctgacgagtt
     1561 tcaccaggat gaaactcttt ccttctctct catccccatt tcatgcaaat aatcattttt
     1621 tattcagtct tacccctatt aaatgtgcat gacacaccag tgaaaccccc attgtgactg
     1681 gccttaacgg tttagtgttt tcttacccaa ttgtagagac tatccacaag gacaatattt
     1741 gtgacttatg ttatgcgcct gctagagttc cggactccgt tcaacttaag gcgaagttgt
     1801 tggcagaaaa tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg gaaatgttct
     1861 atttagaaac aggggaattg cttattaacg aaattgcccc aaggcctcac aactctggac
     1921 attataccat tgatgcttgc gtcacttctc aatttgaagc tcatttgaga tcaatattgg
     1981 atttgccaat gccaaagaat ttcacatctt tctccaccat tacaacgaac gccattatgc
     2041 taaatgttct tggagacaaa catacaaaag ataaagagct agaaacttgc gaaagagcat
     2101 tggcgactcc aggttcctca gtgtacttat atggaaaaga gtctagacct aacagaaaag
     2161 taggtcacat aaatattatt gcctccagta tggcggaatg tgaacaaagg ctgaactaca
     2221 ttacaggtag aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag
     2281 caatggtcaa accattggtt ggaatcatca tgggatcaga ctctgacttg ccggtaatgt
     2341 ctgccgcatg tgcggtttta aaagattttg gcgttccatt tgaagtgaca atagtctctg
     2401 ctcatagaac tccacatagg atgtcagcat atgctatttc cgcaagcaag cgtggaatta
     2461 aaacaattat cgctggagct ggtggggctg ctcacttgcc aggtatggtg gctgcaatga
     2521 caccacttcc tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga gtagattctt
     2581 tacattcaat tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct attaataata
     2641 gtacgaacgc tgcgctgttg gctgtcagac tgcttggcgc ttatgattca agttatacaa
     2701 cgaaaatgga acagttttta ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt
     2761 tagaaactgt cggttacgaa gcttatctag aaaacaagta atatataagt ttattgatat
     2821 acttgtacag caaataatta taaaatgata tacctatttt ttaggctttg ttatgattac
     2881 atcaaatgtg gacttcatac atagaaatca acgcttacag gtgtcctttt ttaagaattt
     2941 catacataag atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt
     3001 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3061 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3121 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3181 cgcataatcg ctgggccatt ctcatgaaga atatcttgaa tttattgtca tattactagt
     3241 tggtgtggaa gtcctaatat cggtgatcaa tatagtggtt gacatgctgg ctagtcaaca
     3301 ttgagccttt tgatcatgca aatatattac ggtattttac aatcaaatat caaacttaac
     3361 tattgacttt ataacttatt taggtggtaa cattcttata aaaaagaaaa aaattactgc
     3421 aaaacagtac tagcttttaa cttgtatcct aggttatcta tgctgtctca ccatagagaa
     3481 tattacctat ttcagaatgt atgtccatga ttcgccgggt aaatacatat aatacacaaa
     3541 tctggcttaa taaagtctat aatatatctc ataaagaagt gctaaattgg ctagtgctat
     3601 atatttttaa gaaaatttct tttgactaag tccatatcga ctttgtaaaa gttcacatta
     3661 gcatacatat attacacgag ccagaaatag taacttttgc ctaaatcaca aattgcaaaa
     3721 tttaattgct tgcaaaaggt cacatgctta taatcaactt ttttaaaaat ttaaaatact
     3781 tttttatttt ttatttttaa acataaatga aataatttat ttattgttta tgattaccga
     3841 aacataaaac ctgctcaaga aaaagaaact gttttgtcct tggaaaaaaa gcactaccta
     3901 ggagcggcca aaatgccgag gctttcatag cttaaactct ttacagaaaa taggcattat
     3961 agatcagttc gagttttctt attcttcctt ccggttttat cgtcacagtt ttacagtaaa
     4021 taagtatcac ctcttagagt tcgatgataa gctgtcaaac atgagaatta attccacatg
     4081 ttaaaatagt gaaggagcat gttcggcaca cagtggaccg aacgtggggt aagtgcacta
     4141 gggtccggtt aaacggatct cgcattgatg aggcaacgct aattatcaac atatagattg
     4201 ttatctatct gcatgaacac gaaatcttta cttgacgact tgaggctgat ggtgtttatg
     4261 caaagaaacc actgtgttta atatgtgtca ctgtttgata ttactgtcag cgtagaagat
     4321 aatagtaaaa gcggttaata agtgtatttg agataagtgt gataaagttt ttacagcgaa
     4381 aagacgataa atacaagaaa atgattacga ggatacggag agaggtatgt acatgtgtat
     4441 ttatatacta agctgccggc ggttgtttgc aagaccgaga aaaggctagc aagaatcggg
     4501 tcattgtagc gtatgcgcct gtgaacattc tcttcaacaa gtttgattcc attgcggtga
     4561 aatggtaaaa gtcaaccccc tgcgatgtat attttcctgt acaatcaatc aaaaagccaa
     4621 atgatttagc attatcttta catcttgtta ttttacagat tttatgttta gatcttttat
     4681 gcttgctttt caaaaggctt gcaggcaagt gcacaaacaa tacttaaata aatactactc
     4741 agtaataacc tatttcttag catttttgac gaaatttgct attttgttag agtcttttac
     4801 accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta atctaagcgc
     4861 atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc tctcggggct
     4921 ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac ctgtcccacc
     4981 tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg cactgagtag
     5041 tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga ggaactcttg
     5101 gtattcttgc cacgactcat ctccatgcag ttggacgatc gatgataagc tgtcaaacat
     5161 gagaattggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     5221 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     5281 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     5341 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     5401 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     5461 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     5521 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     5581 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     5641 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     5701 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     5761 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     5821 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     5881 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     5941 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     6001 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     6061 gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     6121 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     6181 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag
     6241 aataaaaaaa aaatgatgaa ttgaaaagct aattcttgaa gacgaaaggg cctcgtgata
     6301 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7621 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7861 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8161 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8221 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8281 g
//
