LOCUS       40924_2592        3714 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p34S-Km3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Plasposons: modular self-cloning minitransposon derivatives for 
            rapid genetic analysis of gram-negative bacterial genomes
  JOURNAL   Appl. Environ. Microbiol. 64 (7), 2710-2715 (1998)
  PUBMED    9647854
REFERENCE   2  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Improved antibiotic-resistance cassettes through restriction site 
            elimination using Pfu DNA polymerase PCR
  JOURNAL   BioTechniques 25 (5), 772-774 (1998)
  PUBMED    9821576
REFERENCE   3  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-1998) Biotechnology Center for Agriculture and the
            Environment, Cook College, Rutgers University, 59 Dudley Rd., New 
            Brunswick, NJ 08901, USA
REFERENCE   4  (bases 1 to 3714)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3714)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "1998"; volume: "64"; issue: "7"; pages:
            "2710-2715"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "25"; issue: "5"; pages: 
            "772-774"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (12-AUG-1998) Biotechnology Center for Agriculture and the 
            Environment, Cook College, Rutgers University, 59 Dudley Rd., New 
            Brunswick, NJ 08901, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..51
                     /label=multiple cloning site cassette
                     /note="multiple cloning site cassette"
     CDS             162..974
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDEAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    978..1028
                     /label=multiple cloning site cassette
                     /note="multiple cloning site cassette"
     promoter        complement(1034..1052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1071..1087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1095..1111)
                     /label=lac operator
                     /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(1119..1149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1164..1185)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1473..2061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2235..3092)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3093..3197)
                     /label=AmpR promoter
     primer_bind     3671..3687
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3690..3708
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct caaagccacg
       61 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
      121 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
      181 aaacgtcttg ctccaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
      241 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgtttg tatgggaagc
      301 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
      361 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
      421 ttatccggac tcctgatgaa gcatggttac tcaccactgc gatccccggc aaaacagcat
      481 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
      541 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
      601 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
      661 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcac aaacttttgc
      721 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
      781 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
      841 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
      901 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
      961 tcgatgagtt tttctaagag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat
     1021 gcaagcttgt ctccctatag tgagtcgtat tagagcttgg cgtaatcatg gtcatagctg
     1081 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1141 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1201 ctgcccgctt tcgagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1441 aggaaccgta aaaaggccgc gttgctggcg tttttcgata ggctccgccc ccctgacgag
     1501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2521 agtttgcgca acgttgttgg cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2761 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3121 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3181 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3241 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     3301 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3361 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3421 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     3481 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     3541 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3601 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     3661 tcacgacgtt gtaaaacgac ggccagtgaa tttaggtgac actatagaat actc
//
