LOCUS       40924_4924        3473 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pASHSUL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3473)
  AUTHORS   Weeks SD, Drinker M, Loll PJ.
  TITLE     Ligation independent cloning vectors for expression of SUMO fusions
  JOURNAL   Protein Expr. Purif. 53 (1), 40-50 (2007)
  PUBMED    17251035
REFERENCE   2  (bases 1 to 3473)
  AUTHORS   Weeks SD, Drinker M, Loll PJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2007) Biochemistry and Molecular Biology, Drexel 
            University College of Medicine, 245 N 15th Street, Mailstop 497, 
            Philadelphia, PA 19102, USA
REFERENCE   3  (bases 1 to 3473)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3473)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr. Purif."; date: "2007"; volume: "53"; issue: "1"; pages: 
            "40-50"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-JAN-2007) Biochemistry and Molecular Biology, Drexel University 
            College of Medicine, 245 N 15th Street, Mailstop 497, Philadelphia, 
            PA 19102, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..91
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             99..122
                     /codon_start=1
                     /product="TetA fragment"
                     /label=TetA fragment
                     /protein_id="ABN58907.1"
                     /translation="MNSSTKI"
     RBS             130..152
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             165..182
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             189..479
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
     regulatory      505..576
                     /label=lpp terminator region
                     /note="lpp terminator region"
                     /regulatory_class="terminator"
     rep_origin      584..1039
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1066..1170
                     /label=AmpR promoter
     CDS             1171..2028
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             2038..2661
                     /label=TetR
                     /note="tetracycline repressor TetR"
     rep_origin      2817..3405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccatcgaatg gccagatgat taattcctaa tttttgttga cactctatca ttgatagagt
       61 tattttacca ctccctatca gtgatagaga aaagtgaaat gaatagttcg acaaaaatct
      121 agaaataatt ttgtttaact ttaagaagga gatataccat gggtcatcac catcatcatc
      181 acggaggttc tgactccgaa gtcaatcaag aagctaagcc agaggtcaag ccagaagtca
      241 agcctgagac tcacatcaat ttaaaggtgt ccgatggatc ttcagagatc ttcttcaaga
      301 tcaaaaagac cactccttta agaaggctga tggaagcgtt cgctaaaaga cagggtaagg
      361 aaatggactc cttaagattc ttgtacgacg gtattagaat tcaagctgat cagacccctg
      421 aagatttgga catggaggat aacgatatta ttgaggctca ccgcgaacag attggtggcg
      481 tctaaactct cctcgagaag cttgacctgt gaagtgaaaa atggcgcaca ttgtgcgaca
      541 ttttttttgt ctgccgttta ccgctactgc gtcacggatc tccacgcgcc ctgtagcggc
      601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      661 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      781 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      901 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      961 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1021 atattaacgc ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1081 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1141 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1201 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1261 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1321 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1381 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1441 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1501 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1561 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1621 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1681 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1741 ctattaactg gcgaactact tactctagct tcccggcaac aattgataga ctggatggag
     1801 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1861 gataaatctg gagccggtga gcgtggctct cgcggtatca ttgcagcact ggggccagat
     1921 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1981 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta ggaattaatg
     2041 atgtctcgtt tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
     2101 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
     2161 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
     2221 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
     2281 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
     2341 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
     2401 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgcagt ggggcatttt
     2461 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
     2521 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
     2581 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
     2641 cttaaatgtg aaagtgggtc ttaaaagcag cataaccttt ttccgtgatg gtaacttcac
     2701 tagtttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2941 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgacccg aca
//
