LOCUS       sequence-422622-        3701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded mariner transposon with kan selection casette, and modified
            terminal sequences - barcodes can be added via FseI-SbfI restriction
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3701)
  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 3701)
  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_RB-TnV2
FEATURES             Location/Qualifiers
     source          1..3701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     122..144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     136..153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     137..153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      182..570
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            685..794
                     /note="incP origin of transfer"
     CDS             complement(join(958..1749,1750..1818))
                     /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     1581..1600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1819..1923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1995..2013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2632..3426
                     /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(2686..2705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3296..3315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3485..3571
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3663..3690
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctggcggga ctctggggtt cgcgacactg gcagagcatt acgccctgca ggatgcaatg
       61 ggccggccag accggggact tatcagccaa cctgttatgt gttgggtaac gccagggttt
      121 tcccagtcac gacgttgtaa aacgacggcc agtgaattaa ttcttgctta tggaattccc
      181 atgtcagccg ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt
      241 ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa
      301 gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct
      361 gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta
      421 gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag
      481 agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta
      541 tcaacaggtt gaactgctga tcttcagatc ctctacgccg gacgcatcgt ggccggatca
      601 attccgtgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
      661 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
      721 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
      781 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
      841 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
      901 ccccgaagca gggttatgca gcggaaaacg gaattgatcc ccctcgaatt gacgcgttta
      961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1921 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1981 tatcaaccta taaaaatagg cgtatcacga ggccctttcg tcacgcgtcc tcggtaccta
     2041 taacaggttg gctgataagt ccccggtctg ctgatgagac gagcagttac agagatgtta
     2101 cgaacccgtc tcaagcaaca catagatggc gtcgctagta ttaaatgcat atgactttca
     2161 tatagtacca acctggtgat gattcccgga ccaaaacgaa aaaagacgct tttcagcgtc
     2221 ttattgttcg tctttggtac cgagttgaga agagaaaaga aaaccgccga tcctgtccac
     2281 cgcattactg caaggtagtg gacaagaccg gcggtcttaa gttttttggc tgaaagctac
     2341 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcagaggat cgactctaga
     2401 ggatccccgg gtaccgagct cgaattctga ttagcttgca gtgggcttac atggcgatag
     2461 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     2521 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     2581 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2641 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2701 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2761 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     2821 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2881 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2941 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3001 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3061 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3121 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3181 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3241 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3301 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3361 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3421 ttctgagcgg gactctgggg ttcgaaatga ctctagaatg cgagagtagg gaactgccag
     3481 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3541 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     3601 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     3661 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta c
//
