LOCUS       sequence-422620-        3440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mariner transposon with outward-facing pJ23104 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  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 3440)
  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: "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-pOUT
FEATURES             Location/Qualifiers
     source          1..3440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(18..809,810..878))
                     /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     641..660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(879..983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1051..1069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1476..2270
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1530..1549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2140..2159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2329..2415
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2507..2534
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2551..2585
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     rep_origin      2682..3070
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            3185..3294
                     /note="incP origin of transfer"
ORIGIN
        1 ccctcgaatt gacgcgttta ccaatgctta atcagtgagg cacctatctc agcgatctgt
       61 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
      601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
      661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
      721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
      781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
      841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
      901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
      961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1021 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcac
     1081 gcgtcctcgg tacctaacag gttggatgat aagtccccgg tcttcgtatg ccgtcttctg
     1141 cttggcgcgc cctcgagcaa ttgaggaagc ggaacacgta gaaagccagt ccgcagaaac
     1201 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     1261 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     1321 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     1381 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     1441 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     1501 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     1561 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     1621 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     1681 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     1741 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     1801 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     1861 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     1921 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1981 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2041 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2101 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2161 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2221 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     2281 ggggttcgaa atgactctag aatgcgagag tagggaactg ccaggcatca aataaaacga
     2341 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2401 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2461 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2521 acggatggcc tttttgcgtt tctacctgca ttgacagcta gctcagtcct aggtattgtg
     2581 ctagctgcag ggcgcgccaa gcagaagacg gcatacgaag accggggact tatcatccaa
     2641 cctgttagga tccagtgagc gcaacgcaat tggaattccc atgtcagccg ttaagtgttc
     2701 ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt tacatccctg
     2761 gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata ttcttttttt
     2821 tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt aattttattg
     2881 ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc catgagagct
     2941 tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac gttaaacatg
     3001 agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt gaactgctga
     3061 tcttcagatc ctctacgccg gacgcatcgt ggccggatca attccgtgat aggtgggctg
     3121 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     3181 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     3241 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     3301 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     3361 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     3421 gcggaaaacg gaattgatcc
//
