LOCUS       40924_18556        5677 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETHSUL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5677)
  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 5677)
  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 5677)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5677)
  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..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(154..444)
                     /codon_start=1
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(451..468)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(481..503)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(518..542)
                     /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(543..561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        870..947
                     /label=lacI promoter
     CDS             948..2027
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2043..2064
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2839..3027
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3132..3274
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3460..4048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4222..5079)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5080..5184)
                     /label=AmpR promoter
     rep_origin      complement(5211..5666)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcaga agcttctcga ggagagttta gacgccacca atctgttcgc ggtgagcctc
      181 aataatatcg ttatcctcca tgtccaaatc ttcaggggtc tgatcagctt gaattctaat
      241 accgtcgtac aagaatctta aggagtccat ttccttaccc tgtcttttag cgaacgcttc
      301 catcagcctt cttaaaggag tggtcttttt gatcttgaag aagatctctg aagatccatc
      361 ggacaccttt aaattgatgt gagtctcagg cttgacttct ggcttgacct ctggcttagc
      421 ttcttgattg acttcggagt cagaacctcc gtgatgatga tggtgatgac ccatggtata
      481 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
      541 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg
      601 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
      661 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
      721 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
      781 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
      841 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
      901 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
      961 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     1021 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     1081 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     1141 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     1201 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     1261 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     1321 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     1381 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     1441 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     1501 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     1561 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     1621 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     1681 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     1741 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     1801 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     1861 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     1921 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1981 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2041 attaatgtaa gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc
     2101 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2161 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     2221 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     2281 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     2341 aagcaggcca ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg
     2401 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     2461 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     2521 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     2581 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     2641 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     2701 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     2761 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     2821 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     2881 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     2941 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     3001 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3061 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3121 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     3181 actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc
     3241 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     3301 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3361 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3901 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4321 gggcttacca tctggcccca gtgctgcaat gataccgcga gagccacgct caccggctcc
     4381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4441 tttatccgcc tccatccagt ctatcaattg ttgccgggaa gctagagtaa gtagttcgcc
     4501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5161 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa
     5221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5641 gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
