LOCUS       sequence-304579-        8178 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 8178)
  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 8178)
  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 NL60
FEATURES             Location/Qualifiers
     source          1..8178
                     /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(2873..2890)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2873..2889)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2882..2904)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    3646..4808
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3971..4808
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(5143..5946)
                     /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(5947..6162)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6187..6205)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6273..6377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6378..6446,6447..7238)
                     /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(6596..6615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7409..7997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7898..7917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8151..8168
                     /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 tacaggccaa
     1321 attcgctctt agccgtacaa tattactcac cggtgcgatg ccccccatcg taggtgaagg
     1381 tggaaattaa tgatccatct tgagaccaca ggcccacaac agctaccagt ttcctcaagg
     1441 gtccaccaaa aacgtaagcg cttacgtaca tggtcgataa gaaaaggcaa tttgtagatg
     1501 ttaacatcca acgtctcttt caggtcttac ccctattaaa tgtgcatgac acaccagtga
     1561 aacccccatt gtgactggcc ttaacggttt agtgttttct tacccaattg tagagactat
     1621 ccacaaggac aatatttgtg acttatgtta tgcgcctgct agagttccgg actccgttca
     1681 acttaaggcg aagttgttgg cagaaaatgc aatcaaatct tttcccggtt gtggtatatt
     1741 tggtgtggaa atgttctatt tagaaacagg ggaattgctt attaacgaaa ttgccccaag
     1801 gcctcacaac tctggacatt ataccattga tgcttgcgtc acttctcaat ttgaagctca
     1861 tttgagatca atattggatt tgccaatgcc aaagaatttc acatctttct ccaccattac
     1921 aacgaacgcc attatgctaa atgttcttgg agacaaacat acaaaagata aagagctaga
     1981 aacttgcgaa agagcattgg cgactccagg ttcctcagtg tacttatatg gaaaagagtc
     2041 tagacctaac agaaaagtag gtcacataaa tattattgcc tccagtatgg cggaatgtga
     2101 acaaaggctg aactacatta caggtagaac tgatattcca atcaaaatct ctgtcgctca
     2161 aaagttggac ttggaagcaa tggtcaaacc attggttgga atcatcatgg gatcagactc
     2221 tgacttgccg gtaatgtctg ccgcatgtgc ggttttaaaa gattttggcg ttccatttga
     2281 agtgacaata gtctctgctc atagaactcc acataggatg tcagcatatg ctatttccgc
     2341 aagcaagcgt ggaattaaaa caattatcgc tggagctggt ggggctgctc acttgccagg
     2401 tatggtggct gcaatgacac cacttcctgt catcggtgtg cccgtaaaag gttcttgtct
     2461 agatggagta gattctttac attcaattgt gcaaatgcct agaggtgttc cagtagctac
     2521 cgtcgctatt aataatagta cgaacgctgc gctgttggct gtcagactgc ttggcgctta
     2581 tgattcaagt tatacaacga aaatggaaca gtttttatta aagcaagaag aagaagttct
     2641 tgtcaaagca caaaagttag aaactgtcgg ttacgaagct tatctagaaa acaagtaata
     2701 tataagttta ttgatatact tgtacagcaa ataattataa aatgatatac ctatttttta
     2761 ggctttgtta tgattacatc aaatgtggac ttcatacata gaaatcaacg cttacaggtg
     2821 tcctttttta agaatttcat acataagatc cccgggtacc gagctcgaat tcactggccg
     2881 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2941 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3001 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     3061 tgtgcggtat ttcacaccgc ataatcgctg ggccattctc atgaagaata tcttgaattt
     3121 attgtcatat tactagttgg tgtggaagtc ctaatatcgg tgatcaatat agtggttgac
     3181 atgctggcta gtcaacattg agccttttga tcatgcaaat atattacggt attttacaat
     3241 caaatatcaa acttaactat tgactttata acttatttag gtggtaacat tcttataaaa
     3301 aagaaaaaaa ttactgcaaa acagtactag cttttaactt gtatcctagg ttatctatgc
     3361 tgtctcacca tagagaatat tacctatttc agaatgtatg tccatgattc gccgggtaaa
     3421 tacatataat acacaaatct ggcttaataa agtctataat atatctcata aagaagtgct
     3481 aaattggcta gtgctatata tttttaagaa aatttctttt gactaagtcc atatcgactt
     3541 tgtaaaagtt cacattagca tacatatatt acacgagcca gaaatagtaa cttttgccta
     3601 aatcacaaat tgcaaaattt aattgcttgc aaaaggtcac atgcttataa tcaacttttt
     3661 taaaaattta aaatactttt ttatttttta tttttaaaca taaatgaaat aatttattta
     3721 ttgtttatga ttaccgaaac ataaaacctg ctcaagaaaa agaaactgtt ttgtccttgg
     3781 aaaaaaagca ctacctagga gcggccaaaa tgccgaggct ttcatagctt aaactcttta
     3841 cagaaaatag gcattataga tcagttcgag ttttcttatt cttccttccg gttttatcgt
     3901 cacagtttta cagtaaataa gtatcacctc ttagagttcg atgataagct gtcaaacatg
     3961 agaattaatt ccacatgtta aaatagtgaa ggagcatgtt cggcacacag tggaccgaac
     4021 gtggggtaag tgcactaggg tccggttaaa cggatctcgc attgatgagg caacgctaat
     4081 tatcaacata tagattgtta tctatctgca tgaacacgaa atctttactt gacgacttga
     4141 ggctgatggt gtttatgcaa agaaaccact gtgtttaata tgtgtcactg tttgatatta
     4201 ctgtcagcgt agaagataat agtaaaagcg gttaataagt gtatttgaga taagtgtgat
     4261 aaagttttta cagcgaaaag acgataaata caagaaaatg attacgagga tacggagaga
     4321 ggtatgtaca tgtgtattta tatactaagc tgccggcggt tgtttgcaag accgagaaaa
     4381 ggctagcaag aatcgggtca ttgtagcgta tgcgcctgtg aacattctct tcaacaagtt
     4441 tgattccatt gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt ttcctgtaca
     4501 atcaatcaaa aagccaaatg atttagcatt atctttacat cttgttattt tacagatttt
     4561 atgtttagat cttttatgct tgcttttcaa aaggcttgca ggcaagtgca caaacaatac
     4621 ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt
     4681 ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg
     4741 ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa
     4801 tgtaagctct cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa
     4861 agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt
     4921 gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc
     4981 aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg gacgatcgat
     5041 gataagctgt caaacatgag aattgggtaa taactgatat aattaaattg aagctctaat
     5101 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     5161 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     5221 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac
     5281 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc
     5341 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat
     5401 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc
     5461 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc
     5521 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc
     5581 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa
     5641 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt
     5701 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc
     5761 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc
     5821 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt
     5881 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt
     5941 cgacatgatt tatcttcgtt tcctgcatgt ttttgttctg tgcagttggg ttaagaatac
     6001 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
     6061 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
     6121 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac
     6181 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6241 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6361 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6421 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6481 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6541 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6601 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6661 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6721 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6781 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6841 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6901 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6961 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7021 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7081 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7141 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7201 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7261 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7321 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7681 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7741 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7801 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7861 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7921 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7981 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8041 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     8101 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     8161 tgagcgagga agcggaag
//
