LOCUS       sequence-297766-        8783 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 8783)
  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 8783)
  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 2xE
FEATURES             Location/Qualifiers
     source          1..8783
                     /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(3478..3495)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3478..3494)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3487..3509)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    4251..5413
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      4576..5413
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5748..6551)
                     /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(6552..6767)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6792..6810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6878..6982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6983..7051,7052..7843)
                     /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(7201..7220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8014..8602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8503..8522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8756..8773
                     /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 tctcaaatga
     1321 aatcttttat gaaacaatcc ccacagtgga ggggtttcac tttgacgttt ccaagactaa
     1381 gcaaagcatt taattgatac aagttgctgg gatcatttgt acccaaaatc cgtcgacgga
     1441 taagtagtgg gggataagat tcctttgtga taaggttact ttccgccctt acattttcca
     1501 ccttacatgt gtcctctatg tctctttcac aatcaccgac cttatcttct tcttttcatt
     1561 gttgtcgtca gtgcttacgt cttcaagatt cttttcttcg cctggttctt ctttttcaat
     1621 ttctacgtat tcttcttcgt attctggcag tataggatct tgtatctgta cattcttaat
     1681 taaggataag tagtggggga taagattcct ttgtgataag gttactttcc gcccttacat
     1741 tttccacctt acatgtgtcc tctatgtctc tttcacaatc accgacctta tcttcttctt
     1801 ttcattgttg tcgtcagtgc ttacgtcttc aagattcttt tcttcgcctg gttcttcttt
     1861 ttcaatttct acgtattctt cttcgtattc tggcagtata ggatcttgta tctgtacatt
     1921 cttcagaatt tatggcgcgg cgcgggagaa tgcggaggtc gcacggcgga ggcggacgca
     1981 agagatccgg tgaatgaaac gaatcggcct caacgggggt ttcactctgt taccgaggac
     2041 ttggaaacga cgctgacgag tttcaccagg atgaaactct ttccttctct ctcatcccca
     2101 tttcatgcaa ataatcattt tttattcagt cttaccccta ttaaatgtgc atgacacacc
     2161 agtgaaaccc ccattgtgac tggccttaac ggtttagtgt tttcttaccc aattgtagag
     2221 actatccaca aggacaatat ttgtgactta tgttatgcgc ctgctagagt tccggactcc
     2281 gttcaactta aggcgaagtt gttggcagaa aatgcaatca aatcttttcc cggttgtggt
     2341 atatttggtg tggaaatgtt ctatttagaa acaggggaat tgcttattaa cgaaattgcc
     2401 ccaaggcctc acaactctgg acattatacc attgatgctt gcgtcacttc tcaatttgaa
     2461 gctcatttga gatcaatatt ggatttgcca atgccaaaga atttcacatc tttctccacc
     2521 attacaacga acgccattat gctaaatgtt cttggagaca aacatacaaa agataaagag
     2581 ctagaaactt gcgaaagagc attggcgact ccaggttcct cagtgtactt atatggaaaa
     2641 gagtctagac ctaacagaaa agtaggtcac ataaatatta ttgcctccag tatggcggaa
     2701 tgtgaacaaa ggctgaacta cattacaggt agaactgata ttccaatcaa aatctctgtc
     2761 gctcaaaagt tggacttgga agcaatggtc aaaccattgg ttggaatcat catgggatca
     2821 gactctgact tgccggtaat gtctgccgca tgtgcggttt taaaagattt tggcgttcca
     2881 tttgaagtga caatagtctc tgctcataga actccacata ggatgtcagc atatgctatt
     2941 tccgcaagca agcgtggaat taaaacaatt atcgctggag ctggtggggc tgctcacttg
     3001 ccaggtatgg tggctgcaat gacaccactt cctgtcatcg gtgtgcccgt aaaaggttct
     3061 tgtctagatg gagtagattc tttacattca attgtgcaaa tgcctagagg tgttccagta
     3121 gctaccgtcg ctattaataa tagtacgaac gctgcgctgt tggctgtcag actgcttggc
     3181 gcttatgatt caagttatac aacgaaaatg gaacagtttt tattaaagca agaagaagaa
     3241 gttcttgtca aagcacaaaa gttagaaact gtcggttacg aagcttatct agaaaacaag
     3301 taatatataa gtttattgat atacttgtac agcaaataat tataaaatga tatacctatt
     3361 ttttaggctt tgttatgatt acatcaaatg tggacttcat acatagaaat caacgcttac
     3421 aggtgtcctt ttttaagaat ttcatacata agatccccgg gtaccgagct cgaattcact
     3481 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3541 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3601 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     3661 gcatctgtgc ggtatttcac accgcataat cgctgggcca ttctcatgaa gaatatcttg
     3721 aatttattgt catattacta gttggtgtgg aagtcctaat atcggtgatc aatatagtgg
     3781 ttgacatgct ggctagtcaa cattgagcct tttgatcatg caaatatatt acggtatttt
     3841 acaatcaaat atcaaactta actattgact ttataactta tttaggtggt aacattctta
     3901 taaaaaagaa aaaaattact gcaaaacagt actagctttt aacttgtatc ctaggttatc
     3961 tatgctgtct caccatagag aatattacct atttcagaat gtatgtccat gattcgccgg
     4021 gtaaatacat ataatacaca aatctggctt aataaagtct ataatatatc tcataaagaa
     4081 gtgctaaatt ggctagtgct atatattttt aagaaaattt cttttgacta agtccatatc
     4141 gactttgtaa aagttcacat tagcatacat atattacacg agccagaaat agtaactttt
     4201 gcctaaatca caaattgcaa aatttaattg cttgcaaaag gtcacatgct tataatcaac
     4261 ttttttaaaa atttaaaata cttttttatt ttttattttt aaacataaat gaaataattt
     4321 atttattgtt tatgattacc gaaacataaa acctgctcaa gaaaaagaaa ctgttttgtc
     4381 cttggaaaaa aagcactacc taggagcggc caaaatgccg aggctttcat agcttaaact
     4441 ctttacagaa aataggcatt atagatcagt tcgagttttc ttattcttcc ttccggtttt
     4501 atcgtcacag ttttacagta aataagtatc acctcttaga gttcgatgat aagctgtcaa
     4561 acatgagaat taattccaca tgttaaaata gtgaaggagc atgttcggca cacagtggac
     4621 cgaacgtggg gtaagtgcac tagggtccgg ttaaacggat ctcgcattga tgaggcaacg
     4681 ctaattatca acatatagat tgttatctat ctgcatgaac acgaaatctt tacttgacga
     4741 cttgaggctg atggtgttta tgcaaagaaa ccactgtgtt taatatgtgt cactgtttga
     4801 tattactgtc agcgtagaag ataatagtaa aagcggttaa taagtgtatt tgagataagt
     4861 gtgataaagt ttttacagcg aaaagacgat aaatacaaga aaatgattac gaggatacgg
     4921 agagaggtat gtacatgtgt atttatatac taagctgccg gcggttgttt gcaagaccga
     4981 gaaaaggcta gcaagaatcg ggtcattgta gcgtatgcgc ctgtgaacat tctcttcaac
     5041 aagtttgatt ccattgcggt gaaatggtaa aagtcaaccc cctgcgatgt atattttcct
     5101 gtacaatcaa tcaaaaagcc aaatgattta gcattatctt tacatcttgt tattttacag
     5161 attttatgtt tagatctttt atgcttgctt ttcaaaaggc ttgcaggcaa gtgcacaaac
     5221 aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg
     5281 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     5341 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     5401 taaaatgtaa gctctcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     5461 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     5521 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     5581 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     5641 tcgatgataa gctgtcaaac atgagaattg ggtaataact gatataatta aattgaagct
     5701 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     5761 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     5821 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     5881 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     5941 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     6001 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     6061 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     6121 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     6181 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     6241 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     6301 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     6361 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     6421 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     6481 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     6541 gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag
     6601 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     6661 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     6721 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctaattcttg
     6781 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6841 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8761 gtcagtgagc gaggaagcgg aag
//
