LOCUS       40924_18606        5680 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETS2SUL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  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 5680)
  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 5680)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5680)
  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..5680
                     /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(445..468)
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     RBS             complement(484..506)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(521..545)
                     /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(546..564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        873..950
                     /label=lacI promoter
     CDS             951..2030
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2046..2067
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2842..3030
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3135..3277
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3463..4051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4225..5082)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5083..5187)
                     /label=AmpR promoter
     rep_origin      complement(5214..5669)
                     /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 tgatcggctt 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 cagatttttc gaactgcggg tggctccagc tagccatggt
      481 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      541 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
      601 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
      661 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
      721 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
      781 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
      841 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
      901 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
      961 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     1021 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     1081 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     1141 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1201 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1261 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1321 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     1381 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     1441 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     1501 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     1561 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     1621 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     1681 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     1741 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     1801 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     1861 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     1921 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1981 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2041 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     2101 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2161 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     2221 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     2281 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     2341 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     2401 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2461 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2521 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     2581 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     2641 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2701 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2761 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     2821 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     2881 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     2941 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     3001 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3061 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3121 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3181 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     3241 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3301 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3361 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3421 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3481 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3541 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3601 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3901 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagccac gctcaccggc
     4381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4441 aactttatcc gcctccatcc agtctatcaa ttgttgccgg gaagctagag taagtagttc
     4501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt
     5221 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5281 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5341 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5401 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5461 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5521 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5581 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5641 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
