LOCUS       40924_19036        3455 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for bacterial 
            or in vitro expression of an untagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3455)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3455)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and PmeI sites to 
            remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..3455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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)."
     CDS             109..441
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLQGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    454..509
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      573..620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        849..953
                     /label=AmpR promoter
     CDS             954..1811
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(1972..2560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2676..2959
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3189..3275
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3367..3394
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttccccacta gtaataattt tctttaactt tagtaaggag cgatcgccat ggcacaggtt
      121 atcaacacgt ttgacggggt tgcggattat cttcagacat atcataagct acctgataat
      181 tacattacaa aatcagaagc acaagccctc ggctgggtgg catcaaaagg gaaccttgca
      241 gacgtcgctc cggggaaaag catcggcgga gacatcttct caaacaggga aggcaaactc
      301 cagggcaaaa gcggacgaac atggcgtgaa gcggatatta actatacatc aggcttcaga
      361 aattcagacc ggattcttta ctcaagcgac tggctgattt acaaaacaac ggaccattat
      421 cagaccttta caaaaatcag ataatgttta aacgaattcg agctcggtac ccggggatcc
      481 tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
      541 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      601 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggttcgctt gctgtccata
      661 aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg
      721 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
      781 gtttctgcgg actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg
      841 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      901 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      961 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1021 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1081 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1141 gttttccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1201 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1261 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1321 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac
     1381 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1441 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1501 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1561 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1621 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1681 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1741 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1801 ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     1861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2101 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2161 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2401 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2521 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     2581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2641 atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg
     2701 atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg
     2761 ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc
     2821 gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc
     2881 gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     2941 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     3001 ggatcgagat cttttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     3061 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     3121 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     3181 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3241 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     3301 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     3361 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     3421 tttttctaaa tacattcaaa tatgtatccg ctcat
//
