LOCUS       sequence-297757-        8473 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 8473)
  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 8473)
  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 mmPing20B
FEATURES             Location/Qualifiers
     source          1..8473
                     /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(3168..3185)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3168..3184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3177..3199)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3941..5103
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      4266..5103
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5438..6241)
                     /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(6242..6457)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6482..6500)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6568..6672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6673..6741,6742..7533)
                     /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(6891..6910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7704..8292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8193..8212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8446..8463
                     /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 cggaattcgc atgcagtttg acgggtgttg aaataaacaa aaatgaggtg gcggaagaga
     1501 atacgagttt gaggttgggt tagaaacaac aaatgtgtgg gctcatgatg ggttgagttg
     1561 gtgaatgttt tgggctgctc gattgacacc tttgtgagta cgtgttgttg tgcatggctt
     1621 ttggggtcca agcttgagaa tgcagaggtc gcacggcgga ggcggacgca agagatccgg
     1681 tgaatgaaac gaatcggcct caacgggggt ttcgctctgt taccgaggac ttggaaacga
     1741 cgctgacgag tttcaccagg atgaaactct ttccttctct ctcatcccca tttcatgcaa
     1801 ataatcattt tttattcagt cttaccccta ttaaatgtgc atgacacacc agtgaaaccc
     1861 ccattgtgac tggccttaac ggtttagtgt tttcttaccc aattgtagag actatccaca
     1921 aggacaatat ttgtgactta tgttatgcgc ctgctagagt tccggactcc gttcaactta
     1981 aggcgaagtt gttggcagaa aatgcaatca aatcttttcc cggttgtggt atatttggtg
     2041 tggaaatgtt ctatttagaa acaggggaat tgcttattaa cgaaattgcc ccaaggcctc
     2101 acaactctgg acattatacc attgatgctt gcgtcacttc tcaatttgaa gctcatttga
     2161 gatcaatatt ggatttgcca atgccaaaga atttcacatc tttctccacc attacaacga
     2221 acgccattat gctaaatgtt cttggagaca aacatacaaa agataaagag ctagaaactt
     2281 gcgaaagagc attggcgact ccaggttcct cagtgtactt atatggaaaa gagtctagac
     2341 ctaacagaaa agtaggtcac ataaatatta ttgcctccag tatggcggaa tgtgaacaaa
     2401 ggctgaacta cattacaggt agaactgata ttccaatcaa aatctctgtc gctcaaaagt
     2461 tggacttgga agcaatggtc aaaccattgg ttggaatcat catgggatca gactctgact
     2521 tgccggtaat gtctgccgca tgtgcggttt taaaagattt tggcgttcca tttgaagtga
     2581 caatagtctc tgctcataga actccacata ggatgtcagc atatgctatt tccgcaagca
     2641 agcgtggaat taaaacaatt atcgctggag ctggtggggc tgctcacttg ccaggtatgg
     2701 tggctgcaat gacaccactt cctgtcatcg gtgtgcccgt aaaaggttct tgtctagatg
     2761 gagtagattc tttacattca attgtgcaaa tgcctagagg tgttccagta gctaccgtcg
     2821 ctattaataa tagtacgaac gctgcgctgt tggctgtcag actgcttggc gcttatgatt
     2881 caagttatac aacgaaaatg gaacagtttt tattaaagca agaagaagaa gttcttgtca
     2941 aagcacaaaa gttagaaact gtcggttacg aagcttatct agaaaacaag taatatataa
     3001 gtttattgat atacttgtac agcaaataat tataaaatga tatacctatt ttttaggctt
     3061 tgttatgatt acatcaaatg tggacttcat acatagaaat caacgcttac aggtgtcctt
     3121 ttttaagaat ttcatacata agatccccgg gtaccgagct cgaattcact ggccgtcgtt
     3181 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3241 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3301 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3361 ggtatttcac accgcataat cgctgggcca ttctcatgaa gaatatcttg aatttattgt
     3421 catattacta gttggtgtgg aagtcctaat atcggtgatc aatatagtgg ttgacatgct
     3481 ggctagtcaa cattgagcct tttgatcatg caaatatatt acggtatttt acaatcaaat
     3541 atcaaactta actattgact ttataactta tttaggtggt aacattctta taaaaaagaa
     3601 aaaaattact gcaaaacagt actagctttt aacttgtatc ctaggttatc tatgctgtct
     3661 caccatagag aatattacct atttcagaat gtatgtccat gattcgccgg gtaaatacat
     3721 ataatacaca aatctggctt aataaagtct ataatatatc tcataaagaa gtgctaaatt
     3781 ggctagtgct atatattttt aagaaaattt cttttgacta agtccatatc gactttgtaa
     3841 aagttcacat tagcatacat atattacacg agccagaaat agtaactttt gcctaaatca
     3901 caaattgcaa aatttaattg cttgcaaaag gtcacatgct tataatcaac ttttttaaaa
     3961 atttaaaata cttttttatt ttttattttt aaacataaat gaaataattt atttattgtt
     4021 tatgattacc gaaacataaa acctgctcaa gaaaaagaaa ctgttttgtc cttggaaaaa
     4081 aagcactacc taggagcggc caaaatgccg aggctttcat agcttaaact ctttacagaa
     4141 aataggcatt atagatcagt tcgagttttc ttattcttcc ttccggtttt atcgtcacag
     4201 ttttacagta aataagtatc acctcttaga gttcgatgat aagctgtcaa acatgagaat
     4261 taattccaca tgttaaaata gtgaaggagc atgttcggca cacagtggac cgaacgtggg
     4321 gtaagtgcac tagggtccgg ttaaacggat ctcgcattga tgaggcaacg ctaattatca
     4381 acatatagat tgttatctat ctgcatgaac acgaaatctt tacttgacga cttgaggctg
     4441 atggtgttta tgcaaagaaa ccactgtgtt taatatgtgt cactgtttga tattactgtc
     4501 agcgtagaag ataatagtaa aagcggttaa taagtgtatt tgagataagt gtgataaagt
     4561 ttttacagcg aaaagacgat aaatacaaga aaatgattac gaggatacgg agagaggtat
     4621 gtacatgtgt atttatatac taagctgccg gcggttgttt gcaagaccga gaaaaggcta
     4681 gcaagaatcg ggtcattgta gcgtatgcgc ctgtgaacat tctcttcaac aagtttgatt
     4741 ccattgcggt gaaatggtaa aagtcaaccc cctgcgatgt atattttcct gtacaatcaa
     4801 tcaaaaagcc aaatgattta gcattatctt tacatcttgt tattttacag attttatgtt
     4861 tagatctttt atgcttgctt ttcaaaaggc ttgcaggcaa gtgcacaaac aatacttaaa
     4921 taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg ctattttgtt
     4981 agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa taacgccatt
     5041 taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca taaaatgtaa
     5101 gctctcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat ccaaaagttc
     5161 acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc
     5221 tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa ctggcaaacc
     5281 gaggaactct tggtattctt gccacgactc atctccatgc agttggacga tcgatgataa
     5341 gctgtcaaac atgagaattg ggtaataact gatataatta aattgaagct ctaatttgtg
     5401 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     5461 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     5521 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     5581 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     5641 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     5701 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     5761 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     5821 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     5881 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     5941 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     6001 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     6061 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     6121 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     6181 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     6241 tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc
     6301 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     6361 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     6421 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctaattcttg aagacgaaag
     6481 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6541 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6601 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6661 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6721 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6781 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6841 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6901 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6961 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7021 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7081 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7141 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7201 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7261 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7321 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7381 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7441 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7501 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7561 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7621 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7681 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7741 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7801 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7861 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7921 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7981 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8041 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8101 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8161 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8221 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8281 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8341 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8401 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8461 gaggaagcgg aag
//
