LOCUS       pGERM        4807 bp DNA     circular SYN 26-JUL-2024
DEFINITION  PGERM gene disruption vector, complete sequence.
ACCESSION   EF155418
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4807)
  AUTHORS   Shoemaker NB, Wang GR, Salyers AA.
  TITLE     Multiple gene products and sequences required for excision of the 
            mobilizable integrated Bacteroides element NBU1
  JOURNAL   J. Bacteriol. 182 (4), 928-936 (2000)
  PUBMED    10648516
REFERENCE   2  (bases 1 to 4807)
  AUTHORS   Chiang HC, Manchester JK, Gordon JI.
  TITLE     Regulation of nitrogen metabolism by a novel hybrid two-component 
            system protein in a prominent human gut symbiont, Bacteroides 
            thetaiotaomicron
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4807)
  AUTHORS   Chiang HC, Manchester JK, Gordon JI.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-DEC-2006) Center for Genome Sciences, Washington 
            University School of Medicine, 4444 Forest Park Ave. Campus Box 
            8510, Saint Louis, MO 63108, USA
REFERENCE   4  (bases 1 to 4807)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2000"; volume: "182"; issue: "4"; pages: 
            "928-936"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (03-DEC-2006) Center for Genome Sciences, Washington University 
            School of Medicine, 4444 Forest Park Ave. Campus Box 8510, Saint 
            Louis, MO 63108, USA"
FEATURES             Location/Qualifiers
     source          1..4807
                     /mol_type="other DNA"
                     /label=pGERM is pUC19 with the RK2 oriT and ermG
                     /note="pGERM is pUC19 with the RK2 oriT and ermG"
                     /db_xref="taxon:433620"
                     /organism="pGERM gene disruption vector"
     CDS             complement(44..367)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     277..293
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    294..350
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(363..379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    387..403
                     /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(411..441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    456..477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(709..1080)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1113..1224
                     /label=origin of transfer from 56 kb circular plasmid RK2
                     /note="origin of transfer from 56 kb circular plasmid RK2"
     oriT            1113..1222
                     /note="incP origin of transfer"
     rep_origin      complement(1543..2131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(2302..3162)
                     /gene="AmpR"
                     /label=AmpR
     CDS             complement(2302..3162)
                     /codon_start=1
                     /transl_table=11
                     /gene="AmpR"
                     /product="beta lactamase"
                     /label=AmpR
                     /protein_id="ABO69572.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3650..4384
                     /gene="ermG"
                     /label=ermG
     CDS             3650..4384
                     /codon_start=1
                     /transl_table=11
                     /gene="ermG"
                     /product="rRNA methyltransferase"
                     /label=ermG
                     /protein_id="ABO69573.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
ORIGIN
        1 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
       61 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
      121 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
      181 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
      241 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
      301 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
      361 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      421 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
      481 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
      541 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcggtctt gccttgctcg
      601 tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg catggggacg
      661 tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc atggctctgc
      721 cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct cttcgatctt
      781 cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag
      841 ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg
      901 gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta
      961 ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc gttcgtctgg
     1021 aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
     1081 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
     1141 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
     1201 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
     1261 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
     1321 tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc tcttccgctt cctcgctcac
     1381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaatc
     3181 atggccgcgg gattaaaagt cggggattgg tgaacaaaaa ggtgtttctc tctttaagag
     3241 aaatatcgtt ttgctaaaca gttgatattg aggtatcatt ttatcgtaaa agacattttt
     3301 gctcaacaat tgcttgacgg aaatcaacaa attttagcat tttgtaaaaa agtcgctata
     3361 taatttggtg aattggagtt attttcatat ttttgcatcc cgaagagttt ctcttaaaga
     3421 gagaaacatc ttttgcatac cttttccgac cgaattttta tgtcgtaaag aggggctttg
     3481 cagggggtgg actcagaaag atgagaatag atgactattg tagttgaaac acatagaaag
     3541 ttgctgatat acagaccgat acgcatatcg ggatgaacca tgagtacgtt cttttctcaa
     3601 aaaacataaa tattcgaaaa gagatgcaat aaattaagga gaggttataa tgaacaaagt
     3661 aaatataaaa gatagtcaaa attttattac ttcaaaatat cacatagaaa aaataatgaa
     3721 ttgcataagt ttagatgaaa aagataacat ctttgaaata ggtgcaggga aaggtcattt
     3781 tactgctgga ttggtaaaga gatgtaattt tgtaacggcg atagaaattg attctaaatt
     3841 atgtgaggta actcgtaata agctcttaaa ttatcctaac tatcaaatag taaatgatga
     3901 tatactgaaa tttacatttc ctagccacaa tccatataaa atatttggca gcatacctta
     3961 caacataagc acaaatataa ttcgaaaaat tgtttttgaa agttcagcca caataagtta
     4021 tttaatagtg gaatatggtt ttgctaaaat gttattagat acaaacagat cactagcatt
     4081 gctgttaatg gcagaggtag atatttctat attagcaaaa attcctaggt attatttcca
     4141 tccaaaacct aaagtggata gcacattaat tgtattaaaa agaaagccag caaaaatggc
     4201 atttaaagag agaaaaaaat atgaaacttt tgtaatgaaa tgggttaaca aagagtacga
     4261 aaaactgttt acaaaaaatc aatttaataa agctttaaaa catgcgagaa tatatgatat
     4321 aaacaatatt agtttcgaac aatttgtatc gctatttaat agttataaaa tatttaacgg
     4381 ctaaaaacaa taggccacat gcaactgtaa atgtttagtt ataggtaggg tagcataagt
     4441 taaaatgcta ttctgccttt taaaagtatg gtatacattc gtaggcgtcc aataacaagg
     4501 tgtaatcact agtgcggccg ccattattga agcatttatc agggttattg tctcatgagc
     4561 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4621 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4681 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4741 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4801 gcccgtc
//
