LOCUS       sequence-181631-        5910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mariner transposon mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5910)
  AUTHORS   Armbruster CE, Forsyth-DeOrnellas V, Johnson AO, Smith SN, Zhao L, 
            Wu W, Mobley HLT
  TITLE     Genome-wide transposon mutagenesis of Proteus mirabilis: Essential 
            genes, fitness factors for catheter-associated urinary tract 
            infection, and the impact of polymicrobial infection on fitness 
            requirements.
  JOURNAL   PLoS Pathog. 2017 Jun 14;13(6):e1006434. doi: 
            10.1371/journal.ppat.1006434.
  PUBMED    28614382
REFERENCE   2  (bases 1 to 5910)
  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: "PLoS 
            Pathog."; date: "2017-06-14"; volume: "13(6):e1006434. doi"; pages: 
            " 10.1371/journal.ppat.1006434"
COMMENT     Sequence Label: pSAM_AraC
FEATURES             Location/Qualifiers
     source          1..5910
                     /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"
     protein_bind    2632..2653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2668..2673,2674..2691,2692..2698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             complement(2725..3603)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3630..3914
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3837..3856
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5012..5041
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5108..5125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      5152..5540
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            5711..5820
                     /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 gggcgcgcca agcagaagac ggcatacgaa
     2581 gaccggggac ttatcatcca acctgttagg atccagtgag cgcaacgcaa ttaatgtgag
     2641 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2701 tggaattgtg agcggataca tatgttatga caacttgacg gctacatcat tcactttttc
     2761 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     2821 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     2881 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     2941 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     3001 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     3061 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     3121 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     3181 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     3241 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     3301 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     3361 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
     3421 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     3481 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     3541 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     3601 catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt gcatcagaca
     3661 ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa ccccgcttat
     3721 taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt
     3781 ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac tttgctatgc
     3841 catagcattt ttatccataa gattagcgga tcctacctga cgctttttat cgcaactctc
     3901 tactgtttct ccatacccgt tttttgggct aacaggagga attaaccatg gaaaaaaagg
     3961 aatttcgtgt tttgataaaa tactgttttc tgaagggaaa aaatacagtg gaagcaaaaa
     4021 cttggcttga taatgagttt ccggactctg ccccagggaa atcaacaata attgattggt
     4081 atgcaaaatt caagcgtggt gaaatgagca cggaggacgg tgaacgcagt ggacgcccga
     4141 aagaggtggt taccgacgaa aacatcaaaa aaatccacaa aatgattttg aatgaccgta
     4201 aaatgaagtt gatcgagata gcagaggcct taaagatatc aaaggaacgt gttggtcata
     4261 tcattcatca atatttggat atgcggaagc tctgtgcgaa atgggtgccg cgcgagctca
     4321 catttgacca aaaacaacga cgtgttgatg attctaagcg gtgtttgcag ctgttaactc
     4381 gtaatacacc cgagtttttc cgtcgatatg tgacaatgga tgaaacatgg ctccatcact
     4441 acactcctga gtccaatcga cagtcggctg agtggacagc gaccggtgaa ccgtctccga
     4501 agcgtggaaa gactcaaaag tccgctggca aagtaatggc ctctgttttt tgggatgcgc
     4561 atggaataat ttttatcgat tatcttgaga agggaaaaac catcaacagt gactattata
     4621 tggcgttatt ggagcgtttg aaggtcgaaa tcgcggcaaa acggccccac atgaagaaga
     4681 aaaaagtgtt gttccaccaa gacaacgcac cgtgccacaa gtcattgaga acgatggcaa
     4741 aaattcatga attgggcttc gaattgcttc cccacccgcc gtattctcca gatctggccc
     4801 ccagcgactt tttcttgttc tcagacctca aaaggatgct cgcagggaaa aaatttggct
     4861 gcaatgaaga ggtgatcgcc gaaactgagg cctattttga ggcaaaaccg aaggagtact
     4921 accaaaatgg tatcaaaaaa ttggaaggtc gttataatcg ttgtatcgct cttgaaggga
     4981 actatgttga atgaattcga agcttgggcc cgaacaaaaa ctcatctcag aagaggatct
     5041 gaatarmgcc gamgmtyggc cccaacaaaa actcatctca gaagaggatc tgaatagcgc
     5101 cgtcgaccat catcatcatc atcattgagt ttcggtggag gggaattccc atgtcagccg
     5161 ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt
     5221 tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata
     5281 ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt
     5341 aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc
     5401 catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac
     5461 gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt
     5521 gaactgctga tcttcagatc ctctacgccg gacgcatcgt ggccggatca attccgtttt
     5581 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
     5641 tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac
     5701 ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca
     5761 ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg
     5821 ctaccgccgg cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga
     5881 agggcagccc acctatcacg gaattgatcc
//
