LOCUS       sequence-307598-        9276 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 9276)
  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 9276)
  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 4xE
FEATURES             Location/Qualifiers
     source          1..9276
                     /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(3971..3988)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3971..3987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3980..4002)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    4744..5906
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      5069..5906
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(6241..7044)
                     /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(7045..7260)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7285..7303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7371..7475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7476..7544,7545..8336)
                     /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(7694..7713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8507..9095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8996..9015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9249..9266
                     /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 cttcaagctt ggataagtag tgggggataa gattcctttg tgataaggtt actttccgcc
     1981 cttacatttt ccaccttaca tgtgtcctct atgtctcttt cacaatcacc gaccttatct
     2041 tcttcttttc attgttgtcg tcagtgctta cgtcttcaag attcttttct tcgcctggtt
     2101 cttctttttc aatttctacg tattcttctt cgtattctgg cagtatagga tcttgtatct
     2161 gtacattctt aattaaggat aagtagtggg ggataagatt cctttgtgat aaggttactt
     2221 tccgccctta cattttccac cttacatgtg tcctctatgt ctctttcaca atcaccgacc
     2281 ttatcttctt cttttcattg ttgtcgtcag tgcttacgtc ttcaagattc ttttcttcgc
     2341 ctggttcttc tttttcaatt tctacgtatt cttcttcgta ttctggcagt ataggatctt
     2401 gtatctgtac attcttcaga atttatggcg cggcgcggga gaatgcggag gtcgcacggc
     2461 ggaggcggac gcaagagatc cggtgaatga aacgaatcgg cctcaacggg ggtttcactc
     2521 tgttaccgag gacttggaaa cgacgctgac gagtttcacc aggatgaaac tctttccttc
     2581 tctctcatcc ccatttcatg caaataatca ttttttattc agtcttaccc ctattaaatg
     2641 tgcatgacac accagtgaaa cccccattgt gactggcctt aacggtttag tgttttctta
     2701 cccaattgta gagactatcc acaaggacaa tatttgtgac ttatgttatg cgcctgctag
     2761 agttccggac tccgttcaac ttaaggcgaa gttgttggca gaaaatgcaa tcaaatcttt
     2821 tcccggttgt ggtatatttg gtgtggaaat gttctattta gaaacagggg aattgcttat
     2881 taacgaaatt gccccaaggc ctcacaactc tggacattat accattgatg cttgcgtcac
     2941 ttctcaattt gaagctcatt tgagatcaat attggatttg ccaatgccaa agaatttcac
     3001 atctttctcc accattacaa cgaacgccat tatgctaaat gttcttggag acaaacatac
     3061 aaaagataaa gagctagaaa cttgcgaaag agcattggcg actccaggtt cctcagtgta
     3121 cttatatgga aaagagtcta gacctaacag aaaagtaggt cacataaata ttattgcctc
     3181 cagtatggcg gaatgtgaac aaaggctgaa ctacattaca ggtagaactg atattccaat
     3241 caaaatctct gtcgctcaaa agttggactt ggaagcaatg gtcaaaccat tggttggaat
     3301 catcatggga tcagactctg acttgccggt aatgtctgcc gcatgtgcgg ttttaaaaga
     3361 ttttggcgtt ccatttgaag tgacaatagt ctctgctcat agaactccac ataggatgtc
     3421 agcatatgct atttccgcaa gcaagcgtgg aattaaaaca attatcgctg gagctggtgg
     3481 ggctgctcac ttgccaggta tggtggctgc aatgacacca cttcctgtca tcggtgtgcc
     3541 cgtaaaaggt tcttgtctag atggagtaga ttctttacat tcaattgtgc aaatgcctag
     3601 aggtgttcca gtagctaccg tcgctattaa taatagtacg aacgctgcgc tgttggctgt
     3661 cagactgctt ggcgcttatg attcaagtta tacaacgaaa atggaacagt ttttattaaa
     3721 gcaagaagaa gaagttcttg tcaaagcaca aaagttagaa actgtcggtt acgaagctta
     3781 tctagaaaac aagtaatata taagtttatt gatatacttg tacagcaaat aattataaaa
     3841 tgatatacct attttttagg ctttgttatg attacatcaa atgtggactt catacataga
     3901 aatcaacgct tacaggtgtc cttttttaag aatttcatac ataagatccc cgggtaccga
     3961 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4021 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4081 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     4141 attttctcct tacgcatctg tgcggtattt cacaccgcat aatcgctggg ccattctcat
     4201 gaagaatatc ttgaatttat tgtcatatta ctagttggtg tggaagtcct aatatcggtg
     4261 atcaatatag tggttgacat gctggctagt caacattgag ccttttgatc atgcaaatat
     4321 attacggtat tttacaatca aatatcaaac ttaactattg actttataac ttatttaggt
     4381 ggtaacattc ttataaaaaa gaaaaaaatt actgcaaaac agtactagct tttaacttgt
     4441 atcctaggtt atctatgctg tctcaccata gagaatatta cctatttcag aatgtatgtc
     4501 catgattcgc cgggtaaata catataatac acaaatctgg cttaataaag tctataatat
     4561 atctcataaa gaagtgctaa attggctagt gctatatatt tttaagaaaa tttcttttga
     4621 ctaagtccat atcgactttg taaaagttca cattagcata catatattac acgagccaga
     4681 aatagtaact tttgcctaaa tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat
     4741 gcttataatc aactttttta aaaatttaaa atactttttt attttttatt tttaaacata
     4801 aatgaaataa tttatttatt gtttatgatt accgaaacat aaaacctgct caagaaaaag
     4861 aaactgtttt gtccttggaa aaaaagcact acctaggagc ggccaaaatg ccgaggcttt
     4921 catagcttaa actctttaca gaaaataggc attatagatc agttcgagtt ttcttattct
     4981 tccttccggt tttatcgtca cagttttaca gtaaataagt atcacctctt agagttcgat
     5041 gataagctgt caaacatgag aattaattcc acatgttaaa atagtgaagg agcatgttcg
     5101 gcacacagtg gaccgaacgt ggggtaagtg cactagggtc cggttaaacg gatctcgcat
     5161 tgatgaggca acgctaatta tcaacatata gattgttatc tatctgcatg aacacgaaat
     5221 ctttacttga cgacttgagg ctgatggtgt ttatgcaaag aaaccactgt gtttaatatg
     5281 tgtcactgtt tgatattact gtcagcgtag aagataatag taaaagcggt taataagtgt
     5341 atttgagata agtgtgataa agtttttaca gcgaaaagac gataaataca agaaaatgat
     5401 tacgaggata cggagagagg tatgtacatg tgtatttata tactaagctg ccggcggttg
     5461 tttgcaagac cgagaaaagg ctagcaagaa tcgggtcatt gtagcgtatg cgcctgtgaa
     5521 cattctcttc aacaagtttg attccattgc ggtgaaatgg taaaagtcaa ccccctgcga
     5581 tgtatatttt cctgtacaat caatcaaaaa gccaaatgat ttagcattat ctttacatct
     5641 tgttatttta cagattttat gtttagatct tttatgcttg cttttcaaaa ggcttgcagg
     5701 caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt cttagcattt
     5761 ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac acctccgctt
     5821 acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc tggcgtcagt
     5881 ccaccagcta acataaaatg taagctctcg gggctctctt gccttccaac ccagtcagaa
     5941 atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa caagggaata
     6001 aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt tggaaatacg
     6061 agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga ctcatctcca
     6121 tgcagttgga cgatcgatga taagctgtca aacatgagaa ttgggtaata actgatataa
     6181 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
     6241 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
     6301 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
     6361 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
     6421 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
     6481 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
     6541 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat
     6601 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat
     6661 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
     6721 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
     6781 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
     6841 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
     6901 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     6961 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     7021 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcatgttt ttgttctgtg
     7081 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
     7141 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     7201 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     7261 aagctaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7321 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7381 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7441 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7501 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7561 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7621 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7681 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7741 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7801 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7861 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7921 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7981 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     8041 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8101 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8161 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8221 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8281 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8341 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8401 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8461 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8521 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8581 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8641 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8701 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8761 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8821 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8881 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8941 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     9001 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9061 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9121 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     9181 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9241 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
