LOCUS       40924_18501        3476 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for blue/white screening and for inducible bacterial 
            expression of native unfused proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3476)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3476)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Clone into the EcoRV site for optimal spacing from the RBS.
FEATURES             Location/Qualifiers
     source          1..3476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             249..271
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(364..392)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      562..648
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      740..767
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      835..882
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      919..1374
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1492..2349)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2442..3030
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3429..3448)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gacttacaat
       61 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtacg ggcctcttcg
      121 ctattacgcc agcttgcgaa cggtgggtgc gctgcaaggc gattaagttg ggtaacgcca
      181 ggattctccc agtcacgacg ttgtaaaacg acggccagcg agagatcttg attggctagc
      241 agaataattt tgtttaactt taagaaggag atatagatat cgaattcctg cccgggcgtt
      301 gtaatcatag tcataatcaa tactcctgac tgcgttagca atttaactgt gataaactac
      361 cgcattaaag ctattcgatg ataagctgtc aaacatgata attcttgaag acgaaagggc
      421 ctaggctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
      481 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
      541 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      601 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
      661 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
      721 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
      781 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctgagca ataactagca
      841 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      901 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      961 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1021 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1081 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1141 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1201 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1261 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1321 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttctggcg
     1381 gcacgatggc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2341 aatactcata ctcttccttt ttcaatcatg accaaaatcc cttaacgtga gttttcgttc
     2401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2581 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3121 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3181 cgcagcgagt cagtgagcga ggaagccggc gataatggcc tgcttctcgc cgaaacgttt
     3241 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     3301 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     3361 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     3421 gaccggtgaa ttgtgagcgc tcacaattct cgtgacatca taacgtcccg cgaaat
//
