LOCUS       40924_48683        6384 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector RHP-AmpS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6384)
  AUTHORS   Haag AF, Ostermeier C.
  TITLE     Positive Selection Vector for Direct Protein Expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6384)
  AUTHORS   Haag AF, Ostermeier C.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2008) Novartis Institutes for BioMedical Research,
            Novartis Pharma AG, Forum 1, Novartis Campus, Basel CH-4056, 
            Switzerland
REFERENCE   3  (bases 1 to 6384)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6384)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-DEC-2008) Novartis Institutes for BioMedical Research, Novartis 
            Pharma AG, Forum 1, Novartis Campus, Basel CH-4056, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(552..1364)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1486..2074
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2260..2402)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2507..2695)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3470..3491)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3507..4586)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4587..4664)
                     /label=lacI promoter
     promoter        4973..4991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4992..5016
                     /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             5031..5053
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5228..5245
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5249..5272
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             complement(5301..6155)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKH"
     promoter        complement(6156..6227)
                     /label=AmpR promoter
     terminator      6301..6348
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg
      481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      541 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
      601 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
      661 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
      721 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
      781 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
      841 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
      901 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
      961 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1021 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     1081 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1141 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1201 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     1261 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     1321 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     1381 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     1441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1621 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2101 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2161 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2221 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2281 tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct
     2341 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2401 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2461 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2521 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     2581 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     2641 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     2701 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     2761 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     2821 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     2881 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     2941 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3001 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3061 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3121 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3181 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3241 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3301 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3361 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3421 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3481 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3541 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     3601 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     3661 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     3721 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     3781 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     3841 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     3901 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     3961 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4021 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4081 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4141 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4201 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4261 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4321 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4381 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4441 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     4501 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     4561 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     4621 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     4681 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     4741 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     4801 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     4861 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4921 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     4981 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     5041 tttaagaagg agatatacca tggtgaagaa gtaccgttgc aaactctgtg gttacatcta
     5101 cgatcctgaa caaggagatc ccgacagcgg aatcgaaccg gggacaccct tcgaggatct
     5161 tccagacgac tgggtctgtc cgctctgtgg agcttcgaag gaggattttg aaccggttga
     5221 gggatcccat catcatcatc atcacggcct cgaggttctg ttccagggtc ccagagaccg
     5281 cggccgcatt aggtctctca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5401 ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat
     5461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagtaatag
     6241 ctgcaggcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     6301 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     6361 actatatccg gattggcgaa tggg
//
