LOCUS       sequence-422595-        4589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded mariner transposon with crystal violet-inducible outward 
            promoter - 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 4589)
  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 4589)
  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_pJEx
FEATURES             Location/Qualifiers
     source          1..4589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..389
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            504..613
                     /note="incP origin of transfer"
     CDS             complement(join(777..1568,1569..1637))
                     /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     1400..1419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1638..1742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1814..1832
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2227..2235
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3339..4133
                     /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(3393..3412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4003..4022
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4192..4278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4370..4397
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4530..4552
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4544..4561
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4545..4561
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
       61 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
      121 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
      181 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
      241 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
      301 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
      361 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcaa
      421 ttccgtgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca tccgcttgcc
      481 ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac
      541 cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc
      601 acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg
      661 gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac
      721 cccgaagcag ggttatgcag cggaaaacgg aattgatccc cctcgaattg acgcgtttac
      781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1741 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacattt
     1801 atcaacctat aaaaataggc gtatcacgag gccctttcgt cacgcgtcct cggtacctat
     1861 aacaggttgg ctgataagtc cccggtctgc tgagttaaaa gatcttttga attcaaagtt
     1921 ggacacgtgt ccaactttgg gtacatctta aagttggaca cgtgtcctgt caaactattt
     1981 tttgcagttt tttgttatgc agcaacgact catagaaagc acgcagctga tagggtcgac
     2041 gtggtgtttg ctgtgcgttg gacgtgtgga ttgggatgaa aataccccca taaaaaaagg
     2101 cgccgaagcg ccttaatacg ttattagccg atatacgtat actcttactt acttaagagg
     2161 aattgccagc tggggcgccc tctggtaagg ttccccatta atcatccggc tcgtataatg
     2221 tgtggataag gaggtgacaa tatgggctat ctgaatcgcg aagaacgtcg ggaaaccatc
     2281 atgcaggccg ccatgcgcgt cgcgctggat caagggttta ccggtatgac ggtccgcaac
     2341 atcgccaccg ccgcgggcgt cgccgcaggc caggtccacc accatttcac ctcctccggc
     2401 gagctaaaat cccaggcgtt tatccgcgtg atccgcgaga tgatggatct gcagcgcctt
     2461 tcgcgcaccg ccggctggcg tgaacagctg ttttccgccc tcggcagcga ggacggacgg
     2521 ctggagccct atatccgcct gtggcgccag gcgcagctgc tggccgacag cgatccggaa
     2581 atcaaaagcg cgtacctgct gaccatgaac ctgtggcatg acgaagcggt caggatcatt
     2641 cgcgcgggcc atgccgctgg cgaatttacc ctgcgcgata gcgccgaaaa tatcgcctgg
     2701 cggctgattt cgctggtctg tggactggac gggatttacg tgctgggaat gccggaggta
     2761 gacgatgccg cattcacccg ccacctgcag cacgttattc agcttgagtt attttcgtaa
     2821 agcaacacat agatggcgtc gctagtatta aatgcatatg actttcatat agtaccaacc
     2881 tggtgatgat tcccggacca aaacgaaaaa agacgctttt cagcgtctta ttgttcgtct
     2941 ttggtaccga gttgagaaga gaaaagaaaa ccgccgatcc tgtccaccgc attactgcaa
     3001 ggtagtggac aagaccggcg gtcttaagtt ttttggctga aagctactgg gctatctgga
     3061 caagggaaaa cgcaagcgca aagagaaagc agaggatcga ctctagagga tccccgggta
     3121 ccgagctcga attctgatta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     3181 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     3241 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     3301 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3361 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3421 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3481 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3541 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3601 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3661 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3721 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3781 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3841 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3901 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3961 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4021 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4081 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4141 tctggggttc gaaatgactc tagaatgcga gagtagggaa ctgccaggca tcaaataaaa
     4201 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     4261 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     4321 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     4381 ctgacggatg gcctttttgc gtttctacgc tggcgggact ctggggttcg cgacactggc
     4441 agagcattac gccctgcagg atgcaatggg ccggccagac cggggactta tcagccaacc
     4501 tgttatgtgt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4561 tgaattaatt cttgcttatg gaattccca
//
