LOCUS       40924_19861        3425 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for bacterial 
            or in vitro expression of a protein with a C-terminal HQ metal 
            affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3425)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and 
            EcoICRI/Eco53kI sites to remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             70..402
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             430..447
                     /codon_start=1
                     /label=HQ tag
                     /note="HQHQHQ metal affinity tag"
                     /translation="HQHQHQ"
     terminator      543..590
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        819..923
                     /label=AmpR promoter
     CDS             924..1781
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(1942..2530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2646..2929
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     terminator      3159..3245
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3337..3364
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactatagga aattttaact ttactaagga
       61 gcgatcgcca tggcacaggt tatcaacacg tttgacgggg ttgcggatta tcttcagaca
      121 tatcataagc tacctgataa ttacattaca aaatcagaag cacaagccct cggctgggtg
      181 gcatcaaaag ggaaccttgc agacgtcgct ccggggaaaa gcatcggcgg agacatcttc
      241 tcaaacaggg aaggcaaact cccgggcaaa agcggacgaa catggcgtga agcggatatt
      301 aactatacat caggcttcag aaattcagac cggattcttt actcaagcga ctggctgatt
      361 tacaaaacaa cggaccatta tcagaccttt acaaaaatca gataatgttt aatgaccccg
      421 tgtcgagctc atcaacacca acatcaataa tctagagtcg acctgcaggc atgcaagctg
      481 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      541 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      601 gaactatatc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
      661 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
      721 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
      781 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      841 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      901 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
      961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     1141 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1321 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     1681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     1741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     1801 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1861 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2341 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2461 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2521 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2641 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     2701 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     2761 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     2821 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     2881 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     2941 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat cttttcagcc tgatacagat
     3001 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     3061 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     3121 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     3181 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     3241 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     3301 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     3361 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     3421 ctcat
//
