LOCUS       sequence-297768-        8491 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 8491)
  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 8491)
  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 mmPing20F
FEATURES             Location/Qualifiers
     source          1..8491
                     /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(3186..3203)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3186..3202)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3217)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3959..5121
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      4284..5121
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5456..6259)
                     /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(6260..6475)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6500..6518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6586..6690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6691..6759,6760..7551)
                     /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(6909..6928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7722..8310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8211..8230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8464..8481
                     /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 cggaattctt ttgtggtcgt cactgcgttc gtcatacgca ttagtgagtg ggctgtcagg
     1501 acagctcttt tccacgttat tttgttcccc acttgtacta gaggaatctg ctttatcttt
     1561 gcaataaagg caaagatgct tttggtaggt gcgcctaaca attctgcacc attccttttt
     1621 tgtctggtcc ccacaagcca gctgatgaag cttgagaatg cagaggtcgc acggcggagg
     1681 cggacgcaag agatccggtg aatgaaacga atcggcctca acgggggttt cgctctgtta
     1741 ccgaggactt ggaaacgacg ctgacgagtt tcaccaggat gaaactcttt ccttctctct
     1801 catccccatt tcatgcaaat aatcattttt tattcagtct tacccctatt aaatgtgcat
     1861 gacacaccag tgaaaccccc attgtgactg gccttaacgg tttagtgttt tcttacccaa
     1921 ttgtagagac tatccacaag gacaatattt gtgacttatg ttatgcgcct gctagagttc
     1981 cggactccgt tcaacttaag gcgaagttgt tggcagaaaa tgcaatcaaa tcttttcccg
     2041 gttgtggtat atttggtgtg gaaatgttct atttagaaac aggggaattg cttattaacg
     2101 aaattgcccc aaggcctcac aactctggac attataccat tgatgcttgc gtcacttctc
     2161 aatttgaagc tcatttgaga tcaatattgg atttgccaat gccaaagaat ttcacatctt
     2221 tctccaccat tacaacgaac gccattatgc taaatgttct tggagacaaa catacaaaag
     2281 ataaagagct agaaacttgc gaaagagcat tggcgactcc aggttcctca gtgtacttat
     2341 atggaaaaga gtctagacct aacagaaaag taggtcacat aaatattatt gcctccagta
     2401 tggcggaatg tgaacaaagg ctgaactaca ttacaggtag aactgatatt ccaatcaaaa
     2461 tctctgtcgc tcaaaagttg gacttggaag caatggtcaa accattggtt ggaatcatca
     2521 tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg tgcggtttta aaagattttg
     2581 gcgttccatt tgaagtgaca atagtctctg ctcatagaac tccacatagg atgtcagcat
     2641 atgctatttc cgcaagcaag cgtggaatta aaacaattat cgctggagct ggtggggctg
     2701 ctcacttgcc aggtatggtg gctgcaatga caccacttcc tgtcatcggt gtgcccgtaa
     2761 aaggttcttg tctagatgga gtagattctt tacattcaat tgtgcaaatg cctagaggtg
     2821 ttccagtagc taccgtcgct attaataata gtacgaacgc tgcgctgttg gctgtcagac
     2881 tgcttggcgc ttatgattca agttatacaa cgaaaatgga acagttttta ttaaagcaag
     2941 aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt cggttacgaa gcttatctag
     3001 aaaacaagta atatataagt ttattgatat acttgtacag caaataatta taaaatgata
     3061 tacctatttt ttaggctttg ttatgattac atcaaatgtg gacttcatac atagaaatca
     3121 acgcttacag gtgtcctttt ttaagaattt catacataag atccccgggt accgagctcg
     3181 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3241 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3301 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3361 ctccttacgc atctgtgcgg tatttcacac cgcataatcg ctgggccatt ctcatgaaga
     3421 atatcttgaa tttattgtca tattactagt tggtgtggaa gtcctaatat cggtgatcaa
     3481 tatagtggtt gacatgctgg ctagtcaaca ttgagccttt tgatcatgca aatatattac
     3541 ggtattttac aatcaaatat caaacttaac tattgacttt ataacttatt taggtggtaa
     3601 cattcttata aaaaagaaaa aaattactgc aaaacagtac tagcttttaa cttgtatcct
     3661 aggttatcta tgctgtctca ccatagagaa tattacctat ttcagaatgt atgtccatga
     3721 ttcgccgggt aaatacatat aatacacaaa tctggcttaa taaagtctat aatatatctc
     3781 ataaagaagt gctaaattgg ctagtgctat atatttttaa gaaaatttct tttgactaag
     3841 tccatatcga ctttgtaaaa gttcacatta gcatacatat attacacgag ccagaaatag
     3901 taacttttgc ctaaatcaca aattgcaaaa tttaattgct tgcaaaaggt cacatgctta
     3961 taatcaactt ttttaaaaat ttaaaatact tttttatttt ttatttttaa acataaatga
     4021 aataatttat ttattgttta tgattaccga aacataaaac ctgctcaaga aaaagaaact
     4081 gttttgtcct tggaaaaaaa gcactaccta ggagcggcca aaatgccgag gctttcatag
     4141 cttaaactct ttacagaaaa taggcattat agatcagttc gagttttctt attcttcctt
     4201 ccggttttat cgtcacagtt ttacagtaaa taagtatcac ctcttagagt tcgatgataa
     4261 gctgtcaaac atgagaatta attccacatg ttaaaatagt gaaggagcat gttcggcaca
     4321 cagtggaccg aacgtggggt aagtgcacta gggtccggtt aaacggatct cgcattgatg
     4381 aggcaacgct aattatcaac atatagattg ttatctatct gcatgaacac gaaatcttta
     4441 cttgacgact tgaggctgat ggtgtttatg caaagaaacc actgtgttta atatgtgtca
     4501 ctgtttgata ttactgtcag cgtagaagat aatagtaaaa gcggttaata agtgtatttg
     4561 agataagtgt gataaagttt ttacagcgaa aagacgataa atacaagaaa atgattacga
     4621 ggatacggag agaggtatgt acatgtgtat ttatatacta agctgccggc ggttgtttgc
     4681 aagaccgaga aaaggctagc aagaatcggg tcattgtagc gtatgcgcct gtgaacattc
     4741 tcttcaacaa gtttgattcc attgcggtga aatggtaaaa gtcaaccccc tgcgatgtat
     4801 attttcctgt acaatcaatc aaaaagccaa atgatttagc attatcttta catcttgtta
     4861 ttttacagat tttatgttta gatcttttat gcttgctttt caaaaggctt gcaggcaagt
     4921 gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag catttttgac
     4981 gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc
     5041 aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc
     5101 agctaacata aaatgtaagc tctcggggct ctcttgcctt ccaacccagt cagaaatcga
     5161 gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga
     5221 atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct
     5281 tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag
     5341 ttggacgatc gatgataagc tgtcaaacat gagaattggg taataactga tataattaaa
     5401 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     5461 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     5521 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     5581 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     5641 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     5701 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     5761 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     5821 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     5881 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     5941 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     6001 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     6061 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     6121 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     6181 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     6241 tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt
     6301 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     6361 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     6421 aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct
     6481 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7741 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7801 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7861 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7921 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7981 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     8041 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     8101 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8161 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8221 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8281 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8341 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8401 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8461 cgcagcgagt cagtgagcga ggaagcggaa g
//
