LOCUS       pHUE.        5942 bp DNA     circular SYN 28-NOV-2018
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            6xHis-tagged proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pHUE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5942)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        156..174
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    175..199
                     /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             214..236
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             243..245
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             255..272
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             282..299
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             303..530
                     /label=ubiquitin
                     /note="human ubiquitin C"
     misc_feature    528..577
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      636..683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        972..1076
                     /label=AmpR promoter
     CDS             1077..1934
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2108..2696
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2882..3024)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3129..3317)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(4595..4616)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4632..5711)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5712..5789)
                     /label=lacI promoter
ORIGIN
        1 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
       61 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      121 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      181 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      241 ccatgggcag cagccatcat catcatcatc acagcagcgg cctggtgccg cgcggcagcc
      301 atatgcagat ctttgtgaag accctcactg gcaaaaccat cacccttgag gtcgagccca
      361 gtgacaccat tgagaatgtc aaagccaaaa ttcaagacaa ggagggtatc ccacctgacc
      421 agcagcgtct gatatttgcc ggcaaacagc tggaggatgg ccgcactctc tcagactaca
      481 acatccagaa agagtccacc ctgcacctgg tgttgcgcct ccgcggtgga tccgaattcg
      541 agctcggtac cgtcgacgcg gccgcaagct tagatccggc tgctaacaaa gcccgaaagg
      601 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      661 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag
      721 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
      781 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
      841 ctgtgataaa ctaccgcatt aaagctaatt cttgaagacg aaagggcctc gtgatacgcc
      901 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      961 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1021 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1141 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1201 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1261 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1321 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1381 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1441 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1501 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1561 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1621 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1681 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1741 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1801 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1861 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1921 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1981 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2041 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2221 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2761 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2821 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2881 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
     2941 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3001 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3061 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3121 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3181 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     3241 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     3301 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     3361 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     3421 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     3481 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     3541 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     3601 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     3661 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     3721 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     3781 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     3841 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     3901 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     3961 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     4021 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     4081 tgacgatcag cggtccagtg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     4141 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     4201 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     4261 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     4321 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4381 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4441 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4501 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4561 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4621 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4681 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4741 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4801 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4861 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     4921 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4981 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     5041 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5101 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5161 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5221 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5281 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5341 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5401 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5461 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5521 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5581 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5641 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5701 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5761 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5821 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     5881 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     5941 ca
//
