LOCUS       sequence-429337-        3263 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mariner transposon expressing mNeonGreen with kan selection casette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3263)
  AUTHORS   English MA, Alcantar MA, Collins JJ
  TITLE     A self-propagating, barcoded transposon system for the dynamic 
            rewiring of genomic networks.
  JOURNAL   Mol Syst Biol. 2023 Jun 12;19(6):e11398. doi: 
            10.15252/msb.202211398. Epub 2023 Mar 27.
  PUBMED    36970845
REFERENCE   2  (bases 1 to 3263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2023-06-12"; volume: "19(6):e11398. doi"; pages: " 
            10.15252/msb.202211398. Epub 2023 Mar 27"
COMMENT     Sequence Label: pDonor_Tn[GFP]
FEATURES             Location/Qualifiers
     source          1..3263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            604..713
                     /note="incP origin of transfer"
     primer_bind     893..911
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1269..1288)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1280..2095
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1280..2095
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1353..1372)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2123..2157
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             2243..2953
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2498..2507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      2978..3064
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3156..3183
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctctagagta gtagtccatg ggtatggaca gcggtggagg ggaattccca tgtcagccgt
       61 taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt
      121 acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat
      181 tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta
      241 attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc
      301 atgagagctt agtacgttag ccatgagcat gagagcttag tacgttagcc atgagagctt
      361 agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacatga
      421 gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg aactgctgat
      481 cttcagatcc tctacgccgg acgcatcgtg gccggatcaa ttccgtgata ggtgggctgc
      541 ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg
      601 tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
      661 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
      721 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
      781 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
      841 cgtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      901 tatcacgagg ccctttcgtc acgcgtcctc ggtacctaac aggttggatg ataagtcccc
      961 ggtcttcgta tgccgtcttc tgcttggcgc gccctcgagc aattgccgac cgaattttta
     1021 tgtcgtaaag aggggctttg cagggggtgg actcagaaag atgagaatag atgactattg
     1081 tagttgaaac acatagaaag ttgctgatat acagaccgat acgcatatcg gctgatcctt
     1141 caactcagca aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg
     1201 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1261 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg
     1321 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1381 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     1441 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1501 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1561 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     1621 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1681 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc
     1741 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1801 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     1861 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1921 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1981 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2041 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2101 attggttaat tggttgtaac acttgacagc tagctcagtc ctaggtataa tgctagcggg
     2161 cccaagttca cttaaaaagg agatcaacaa tgaaagcaat tttcgtactg aaacatctta
     2221 atcatgcgcc ggaggttttc taatggtgag caagggcgag gaggataaca tggcctctct
     2281 cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt
     2341 gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa
     2401 gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca
     2461 gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg
     2521 ataccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg
     2581 ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc
     2641 tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa
     2701 gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa
     2761 tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc
     2821 taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa
     2881 gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga
     2941 gctgtacaag taacgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     3001 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     3061 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     3121 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     3181 ttttgcgttt ctacctgcag ggcgcgccaa gcagaagacg gcatacgaag accggggact
     3241 tatcatccaa cctgttagga tcc
//
