LOCUS       40924_25191        6777 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Ubiquitin protease vector pHUsp2-cc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6777)
  AUTHORS   Catanzariti AM, Soboleva TA, Jans DA, Board PG, Baker RT.
  TITLE     An efficient system for high-level expression and easy purification 
            of authentic recombinant proteins
  JOURNAL   Protein Sci. 13 (5), 1331-1339 (2004)
  PUBMED    15096636
REFERENCE   2  (bases 1 to 6777)
  AUTHORS   Catanzariti AM, Baker RT.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-SEP-2004) John Curtin School of Medical Research, 
            Australian National University, Mills Road, Canberra, ACT 2601, 
            Australia
REFERENCE   3  (bases 1 to 6777)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6777)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2004"; volume: "13"; issue: "5"; pages: "1331-1339"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-SEP-2004) John Curtin School of Medical Research, Australian 
            National University, Mills Road, Canberra, ACT 2601, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6777
                     /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)."
     RBS             79..101
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             120..137
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             147..164
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     terminator      1309..1356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1531..1559)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        1672..1776
                     /label=AmpR promoter
     CDS             1777..2634
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2808..3396
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3582..3724)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3829..4017)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5295..5316)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5332..6411)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6412..6489)
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
      121 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg ctcgaggatc
      181 ctctgctcac caaagccaag aattcaaaga gtgcccaggg tctggctggt cttcgaaacc
      241 ttgggaacac gtgcttcatg aactcaattc ttcagtgcct gagcaacacc cgagagctga
      301 gagattactg cctccagagg ctgtacatgc gggacctcgg ccacaccagc agcgctcaca
      361 cggccctcat ggaagagttt gcaaaactaa tccagaccat atggacgtcg tcccccaatg
      421 atgtggtgag cccatctgag ttcaagaccc agatccagag atatgcgcca cgcttcatgg
      481 gctataatca gcaggatgct caggaattcc ttcgtttcct tctggatggt ctccacaatg
      541 aggtgaaccg ggtggcagca aggcctaagg ccagccctga gacccttgat catctccctg
      601 atgaagaaaa ggggcgacag atgtggagga agtatctgga aagggaagac agtcggattg
      661 gggatctctt cgttgggcag ctgaagagct ccctcacatg caccgattgt ggctactgct
      721 ctacagtctt cgatcccttc tgggatctct cgttgcccat cgcaaagaga ggttaccctg
      781 aggtgacgtt aatggattgt atgaggctct tcaccaaaga ggacatattg gatggtgatg
      841 agaagccaac ttgctgccgc tgccgagcca gaaaacgatg cataaaaaag ttctctgtcc
      901 agaggttccc aaagatcttg gtgctccacc tgaagcgatt ctcagaatcc aggatacgaa
      961 ccagcaagct cacaacattt gtgaatttcc cactaagaga cctggacttg agagaatttg
     1021 cttcagaaaa caccaaccat gctgtttaca acctgtatgc tgtgtccaat cactccggaa
     1081 ccaccatggg aggccactat acagcctact gccgaagtcc ggttacaggc gaatggcaca
     1141 ctttcaatga ttccagtgtc acacccatgt cctccagcca agtgcgcacc agcgacgcct
     1201 atttgctctt ctatgaactg gccagtccac cctcccgtat gtaaggatcc ggctgctaac
     1261 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1321 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1381 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
     1441 tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc
     1501 gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa
     1561 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1621 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1981 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2281 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2641 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2941 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3481 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3541 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3601 cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct
     3661 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3721 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3781 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3841 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3901 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3961 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     4021 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     4081 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4141 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4201 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4261 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4321 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     4381 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     4441 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     4501 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     4561 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     4621 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     4681 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     4741 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccagtg
     4801 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     4861 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     4921 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     4981 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     5041 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     5101 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     5161 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     5221 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5281 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     5341 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5401 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     5461 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5521 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5581 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     5641 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5701 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5761 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5821 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5881 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5941 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     6001 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     6061 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     6121 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     6181 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6241 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6301 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     6361 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     6421 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6481 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     6541 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     6601 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6661 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6721 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcg
//
