LOCUS       sequence-297728-        7112 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 7112)
  AUTHORS   Payero L, Outten G, Burckhalter CE, Hancock, CN
  TITLE     Alteration of the Ping and Pong ORF1 proteins allows for hyperactive
            transposition of mPing
  JOURNAL   Journal of the South Carolina Academy of Science 2016, 14 (2):1-6.
REFERENCE   2  (bases 1 to 7112)
  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: "Journal of 
            the South Carolina Academy of Science 2016, 14 (2):1-6."
COMMENT     Sequence Label: pAG413 GAL ORF1 Shuffle1 NLS
FEATURES             Location/Qualifiers
     source          1..7112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    76..579
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2798..3239
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2798..2915
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3210..3233
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3254..3270
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3254..3270
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3278..3302
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4704..4728)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4761..4777)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4762..4778)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4780..5027
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4796..4814)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5045..5064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5046..5064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5074..5091)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5074..5090)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5083..5105)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5231..5686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5318..5337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5528..5549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5946..6608)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6609..6796)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6943..6962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7062..7084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
      121 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
      181 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
      241 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
      301 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
      361 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
      421 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
      481 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
      541 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagc tctagtacgg attagaagcc gccgagcggg
     2821 tgacagccct ccgaaggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc
     2881 tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag
     2941 cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatga
     3001 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
     3061 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atgcaaaaac
     3121 tgcataacca ctttaactaa tactttcaac attttcggtt tgtattactt cttattcaaa
     3181 tgtaataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
     3241 aaaaaccccg gattctagaa ctagtggatc ccccatcaca agtttgtaca aaaaagcagg
     3301 cttcatggat ccgtcgccgg ccgtggatcc gtcgccggcc gtggatccgt cgccggctgc
     3361 tgaaacccgg cggcgtgcaa ccgggaaagg aggcaaacag cgcgggggca agcaactagg
     3421 attgaagagg ccgccgccga tttctgtccc ggccaccccg cctcctgctg cgacgtcttc
     3481 atcccctgct gcgccgacgg ccatcccacc acgaccaccg caatcttcgc cgattttcgt
     3541 ccccgattcg ccgaatccgt caccggctgc gccgacctcc tctcttgctt cggggacatc
     3601 gacggcaagg ccaccgcaac cacaaggagg aggatgggga ccaacatcga ccatttcccc
     3661 aaactttgca tctttctttg gaaaccaaca agacccaaat tcatgtttgg tcaggggtta
     3721 tcctccagga gggtttgtca attttattca acaaaattgt ccgccgcagc cacaacagca
     3781 aggtgaaaat tttcatttcg ttggtcacaa tatggggttc aacccaatat ctccacagcc
     3841 accaagtgcc tacggaacac caacacccca agctacgaac caaggcactt caacaaacat
     3901 tatgattgat gaagaggaca acaatgatga cagtagggca gcaaagaaaa gatggactca
     3961 tgaagaggaa gagagactgg ccagtgcttg gttgaatgct tctaaagact caattcatgg
     4021 gaatgataag aaaggtgata cattttggaa ggaagtcact gatgaattta acaagaaagg
     4081 gaatggaaaa cgtaggaggg aaattaacca actgaaggtt cactggtcaa ggttgaagtc
     4141 agcgatctct gagttcaatg actattggag tacggttact caaatgcata caagcggata
     4201 ctcagacgac atgcttgaga aagaggcaca gaggctgtat gcaaacaggt ttggaaaacc
     4261 ttttgcgttg gtccattggt ggaagatact caaaagagag cccaaatggt gtgctcagtt
     4321 tgaaaagagg aaaaggaaga gcgaaatgga tgctgttcca gaacagcaga aacgtcctat
     4381 tggtagagaa gcagcaaagt ctgagcgcaa aagaaagcgc aagaaagaaa atgttatgga
     4441 aggcattgtc ctcctagggg acaatgtcca gaaaattatc aaagtgacgc aagatcggaa
     4501 gctggagcgt gagaaggtca ctgaagcaca gattcacatt tcaaacgtaa atttgaaggc
     4561 agcagaacag caaaaagaag caaagatgtt tgaggtatac aattccctgc tcactcaaga
     4621 tacaagtaac atgtctgaag aacagaaggc tcgccgagac aaggcattac aaaagctgga
     4681 ggaaaagtta tttgctgact aagacccagc tttcttgtac aaagtggtga tgggctgcag
     4741 gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta gttatgtcac
     4801 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     4861 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     4921 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     4981 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa
     5041 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     5101 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5161 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5221 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5641 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     5701 ttctccttac gcatctgtgc ggtatttcac accgcataga tccgtcgagt tcaagagaaa
     5761 aaaaaagaaa aagcaaaaag aaaaaaggaa agcgcgcctc gttcagaatg acacgtatag
     5821 aatgatgcat taccttgtca tcttcagtat catactgttc gtatacatac ttactgacat
     5881 tcataggtat acatatatac acatgtatat atatcgtatg ctgcagcttt aaataatcgg
     5941 tgtcactaca taagaacacc tttggtggag ggaacatcgt tggtaccatt gggcgaggtg
     6001 gcttctctta tggcaaccgc aagagccttg aacgcactct cactacggtg atgatcattc
     6061 ttgcctcgca gacaatcaac gtggagggta attctgctag cctctgcaaa gctttcaaga
     6121 aaatgcggga tcatctcgca agagagatct cctactttct ccctttgcaa accaagttcg
     6181 acaactgcgt acggcctgtt cgaaagatct accaccgctc tggaaagtgc ctcatccaaa
     6241 ggcgcaaatc ctgatccaaa cctttttact ccacgcacgg cccctagggc ctctttaaaa
     6301 gcttgaccga gagcaatccc gcagtcttca gtggtgtgat ggtcgtctat gtgtaagtca
     6361 ccaatgcact caacgattag cgaccagccg gaatgcttgg ccagagcatg tatcatatgg
     6421 tccagaaacc ctatacctgt gtggacgtta atcacttgcg attgtgtggc ctgttctgct
     6481 actgcttctg cctctttttc tgggaagatc gagtgctcta tcgctagggg accacccttt
     6541 aaagagatcg caatctgaat cttggtttca tttgtaatac gctttactag ggctttctgc
     6601 tctgtcatct ttgccttcgt ttatcttgcc tgctcatttt ttagtatatt cttcgaagaa
     6661 atcacattac tttatataat gtataattca ttatgtgata atgccaatcg ctaagaaaaa
     6721 aaaagagtca tccgctaggt ggaaaaaaaa aaatgaaaat cattaccgag gcataaaaaa
     6781 atatagagtg tactagagga ggccaagagt aatagaaaaa gaaaattgcg ggaaaggact
     6841 gtgttatgac ttccctgact aatgccgtgt tcaaacgata cctggcagtg actcctagcg
     6901 ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc agtacaatct gctctgatgc
     6961 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     7021 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     7081 gaggttttca ccgtcatcac cgaaacgcgc ga
//
