LOCUS       sequence-194287-        4263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transposon plasmid, compatible with Tn-Seq.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4263)
  AUTHORS   Lee HH, Ostrov N, Wong BG, Gold MA, Khalil AS, Church GM
  TITLE     Functional genomics of the rapidly replicating bacterium Vibrio 
            natriegens by CRISPRi.
  JOURNAL   Nat Microbiol. 2019 Apr 8. pii: 10.1038/s41564-019-0423-8. doi: 
            10.1038/s41564-019-0423-8.
  PUBMED    30962569
REFERENCE   2  (bases 1 to 4263)
  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: "Nat 
            Microbiol. 2019 Apr 8. pii: 10.1038/s41564-019-0423-8. doi: 
            10.1038/s41564-019-0423-8."
COMMENT     Sequence Label: pMarC9-R6k
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1286..1305)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1297..2112
                     /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"
     misc_feature    2199..2255
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     rep_origin      2334..2722
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            2741..2849
                     /note="incP origin of transfer"
     terminator      complement(2996..3035)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(3039..3830,3831..3899))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3900..3991)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtggggaagc aattcgaagc ccaattcatg aatttttgcc atcgttctca atgacttgtg
       61 gcacggtgcg ttgtcttggt ggaacaacac ttttttcttc ttcatgtggg gccgttttgc
      121 cgcgatttcg accttcaaac gctccaataa cgccatataa tagtcactgt tgatggtttt
      181 tcccttctca agataatcga taaaaattat tccatgcgca tcccaaaaaa cagaggccat
      241 tactttgcca gcggactttt gagtctttcc acgcttcgga gacggttcac cggtcgctgt
      301 ccactcagcc gactgtcgat tggactcagg agtgtagtga tggagccatg tttcatccat
      361 tgtcacatat cgacggaaaa actcgggtgt attacgagtt aacagctgca aacaccgctt
      421 agaatcatca acacgtcgtt gtttttggtc aaatgtgagc tcgcgcggca cccatttcgc
      481 acagagcttc cgcatatcca aatattgatg aatgatatga ccaacacgtt cctttgatat
      541 ctttaaggcc tctgctatct cgatcaactt cattttacgg tcattcaaaa tcattttgtg
      601 gatttttttg atgttttcgt cggtaaccac ctctttcggg cgtccactgc gttcaccgtc
      661 ctccgtgctc atttcaccac gcttgaattt tgcataccaa tcaattattg ttgatttccc
      721 tggggcagag tccggaaact cattatcaag ccaagttttt gcttccactg tattttttcc
      781 cttcagaaaa cagtatttta tcaaaacacg aaattccttt ttttccatat gtttcacaat
      841 aacaaaagtt gcttcacaaa agacgctcta tctcacaaac taattgactt acagacgtca
      901 aattttgaca cgaatcattt gaaggttggt actatataaa aataatatgc atctaatact
      961 agcgacgcca tctatgtgtc atccatcaca ctggcggccg ctctacgggg tctgacgctc
     1021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1141 cttggtctga cagcagctgt aacaggttgg atgataagtc cccggtctag acgtcttgtg
     1201 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1261 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca gcgtgaaacg
     1321 agctgtagcc gtccgcgtct gaacagcaac atggatgcgg atctgtatgg ctataaatgg
     1381 gcgcgtgata acgtgggtca gagcggcgcg accatttatc gtctgtatgg caaaccggat
     1441 gcgccggaac tgtttctgaa acatggcaaa ggcagcgtgg cgaacgatgt gaccgatgaa
     1501 atggtgcgtc tgaactggct gaccgaattt atgccgctgc cgaccattaa acattttatt
     1561 cgcaccccgg atgatgcgtg gctgctgacc accgcgattc cgggcaaaac cgcgtttcag
     1621 gtgctggaag aatatccgga tagcggcgaa aacattgtgg atgcgctggc cgtgtttctg
     1681 cgtcgtctgc atagcattcc ggtgtgcaac tgcccgttta acagcgatcg tgtgtttcgt
     1741 ctggcccagg cgcagagccg tatgaacaac ggcctggtgg atgcgagcga ttttgatgat
     1801 gaacgtaacg gctggccggt ggaacaggtg tggaaagaaa tgcataaact gctgccgttt
     1861 agcccggata gcgtggtgac ccacggcgat tttagcctgg ataacctgat tttcgatgaa
     1921 ggcaaactga ttggctgcat tgatgtgggc cgtgtgggca ttgcggatcg ttatcaggat
     1981 ctggccattc tgtggaactg cctgggcgaa tttagcccga gcctgcaaaa acgtctgttt
     2041 cagaaatatg gcattgataa tccggatatg aacaaactgc aatttcatct gatgctggat
     2101 gaatttttct aataattaat tggggaccct agaggtcccc ttttttattt taaaaatttt
     2161 ttcacaaaac ggtttacaag cataactagt gcggccgcaa gcttgcatgc ctgcaggtcg
     2221 actctagagg atccccgggt accgagctcg aattcgcgcg gccgcggcct aggcggcctt
     2281 aattaataga ccggggactt atcatccaac ctgttacagc tgggcgcgcc ccatgtcagc
     2341 cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc
     2401 gttacatccc tggcttgttg tccacaaccg ttaaacctta aaggctttaa aagccttata
     2461 tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg ctgatttata
     2521 ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtta
     2581 gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg agagcttagt
     2641 acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac tatcaacagg
     2701 ttgaactgct gatcttcaga tcggcgcgcc ggccggccta cggccagcct cgcagagcag
     2761 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     2821 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     2881 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     2941 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaaggacaag ggtcgtccaa
     3001 aaaaaaaggc tccaaaagga gcctttaatt gtatcggttt accaatgttt aatcaggctc
     3061 gcgccaattt ccgcaatctg acggttacgt tcatccatcg tcgcctggct gccggtggta
     3121 taaatcacca caatacggct cggtttacca tccgggccca gcgccgcaat aatgccacgg
     3181 ctgccacgtt cgcccgcacc gcttttatcc gcaataaacc agccagccgg cagcgcgcta
     3241 cgcagcagcg gacccgccac tttatccgct tccatccaat caatcagttg ctggcggctt
     3301 gccagggtca gcagctcacc cgtcagcagt ttacgcaggg tggtcgccat tgctgccggc
     3361 atggtggtat cacgttcatc gttcggaatc gcttcgttca gttccggttc ccaacgatcc
     3421 agacgggtca catgatcgcc catgttatgc agaaacgcgg tcagttcttt cggaccgcca
     3481 atggtcgtca gcagcaggtt cgccgcggtg ttatcgctca tggtaatcgc cgcgctgcac
     3541 agttcacgca cggtcatgcc atcggtcaga tgtttttcgg tcaccgggct atattccacc
     3601 agatcgttct ggctataatg aatacgacgg cccagttgtt cctggcccgc atccacacgg
     3661 ctcagaaccg caccgcacag cagcacttta aaggtgctca tcatcggaaa acgttcttcc
     3721 ggacgaaaag attccagaat tttgccgctg ttcagatcca gttcaatata gcccacgcgc
     3781 gcacccagtt gatcttccgc atctttcact ttcaccaggg tttccggatg cgcaaacacc
     3841 ggcaggcaaa acgccgcaaa aaacggaatc agcgccacac gaaaatgctg aatgctcata
     3901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3961 atatttgaat gtatttagaa aaataaacaa ataggggaat taaaaaaaag cccgctcatt
     4021 aggcgggcta ctaatttaaa tcgattattc aacatagttc ccttcaagag cgatacaacg
     4081 attataacga ccttccaatt ttttgatacc attttggtag tactccttcg gttttgcctc
     4141 aaaataggcc tcagtttcgg cgatcacctc ttcattgcag ccaaattttt tccctgcgag
     4201 catccttttg aggtctgaga acaagaaaaa gtcgctgggg gccagatctg gagaatacgg
     4261 cgg
//
