LOCUS       sequence-412227-        3588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector for transposome-mediated saturation mutagenesis and Tn-Seq in
            S. aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3588)
  AUTHORS   Lo H-Y, Long DR, Holmes EA, Penewit K, Hodgson T, Lewis JD, Waalkes 
            A, Salipante SJ
  TITLE     Transposon sequencing identifies genes impacting Staphylococcus 
            aureus invasion in a human macrophage model.
  JOURNAL   Infect Immun. 2023 Oct 17;91(10):e0022823. doi: 
            10.1128/iai.00228-23. Epub 2023 Sep 7.
  PUBMED    37676013
REFERENCE   2  (bases 1 to 3588)
  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: "Infect 
            Immun."; date: "2023-10-17"; volume: "91(10):e0022823. doi"; pages: 
            " 10.1128/iai.00228-23. Epub 2023 Sep 7"
COMMENT     Sequence Label: pAureus-TnCAM
FEATURES             Location/Qualifiers
     source          1..3588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    244..262
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     misc_feature    1177..1198
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_feature    1233..1253
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     misc_feature    1327..1345
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(1367..1383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1380..1402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1391..1407
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1415..1421,1422..1439,1440..1445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1460..1481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1598..1615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1769..2357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1849..1868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2528..3319,3320..3388))
                     /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     3151..3170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3389..3493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3561..3579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 cagctgtctc ttatacacat ctgggcccgg atccgtcgac tcatgacctg caggaaacac
      301 ttttttgttt acttctcatt tttaattagt tataattaac taaataatag agcattaaat
      361 atatttaata aaacttattt aatgcaaaat tatgactaac atatctataa taaataaaga
      421 ttagatatca atatattatc gggcaaatgt atcgagcaag atgcatgcat caaattagga
      481 ggatgattat ttatgaactt taataaaatt gatttagaca attggaagag aaaagagata
      541 tttaatcatt atttgaacca acaaacgact tttagtataa ccacagaaat tgatattagt
      601 gttttatacc gaaacataaa acaagaagga tataaatttt accctgcatt tattttctta
      661 gtgacaaggg tgataaactc aaatacagct tttagaactg gttacaatag cgacggagag
      721 ttaggttatt gggataagtt agagccactt tatacaattt ttgatggtgt atctaaaaca
      781 ttctctggta tttggactcc tgtaaagaat gacttcaaag agttttatga tttatacctt
      841 tctgatgtag agaaatataa tggttcgggg aaattgtttc ccaaaacacc tatacctgaa
      901 aatgcttttt ctctttctat tattccatgg acttcattta ctgggtttaa cttaaatatc
      961 aataataata gtaattacct tctacccatt attacagcag gaaaattcat taataaaggt
     1021 aattcaatat atttaccgct atctttacag gtacatcatt ctgtttgtga tggttatcat
     1081 gcaggattgt ttatgaactc tattcaggaa ttgtcagata ggcctaatga ctggctttta
     1141 taatgataat cccgggctcg aggtcgacgt ttcttcgaat tcgcggccgc ttctagagcg
     1201 caaaaaaccc cgcttcggcg gggttttttc gctactagta gcggccgctg caggaagaaa
     1261 cagtcgacct gcaggcatgc aacgttgcca acgactacgc actagccaac aagagcttca
     1321 gggttgagat gtgtataaga gacagctgaa gcttggcgta atcatggtca tagctgtttc
     1381 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1441 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3541 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
