LOCUS       40924_18356        7182 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET21b+PA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7182)
  AUTHORS   Vahedi F, Sankian M, Mahmoudi M.
  TITLE     Functional expression of Bacillus anthracis protective antigen in E.
            coli
  JOURNAL   Appl. Biochem. Biotechnol. 157 (3), 554-561 (2009)
  PUBMED    18654743
REFERENCE   2  (bases 1 to 7182)
  AUTHORS   Vahedi F, Mahmoudi M.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2007) Immunology, Razi Vaccine and Serum 
            Institute, Ahmad Abad, Mashhad 91735, Iran
REFERENCE   3  (bases 1 to 7182)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7182)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Biochem. Biotechnol."; date: "2009"; volume: "157"; issue: "3"; 
            pages: "554-561"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-APR-2007) Immunology, Razi Vaccine and Serum Institute, Ahmad 
            Abad, Mashhad 91735, Iran"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7182
                     /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(140..157)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(206..238)
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     RBS             complement(246..268)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(283..307)
                     /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(308..326)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        635..712
                     /label=lacI promoter
     CDS             713..1792
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1808..1829
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2604..2792
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2897..3039
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3225..3813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3987..4844)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4845..4949)
                     /label=AmpR promoter
     rep_origin      complement(4976..5431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     gene            5462..7147
                     /gene="pa"
                     /label=pa
                     /note="protective antigen fragment; derived from Bacillus 
                     anthracis isolate 34F2 plasmid pXO1"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat
      241 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
      301 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
      361 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      421 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      481 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
      541 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
      601 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
      661 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
      721 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
      781 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
      841 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
      901 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
      961 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     1021 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     1081 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     1141 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     1201 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     1261 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     1321 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     1381 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     1441 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     1501 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     1561 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     1621 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     1681 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1741 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1801 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     1861 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     1921 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     1981 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     2041 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     2101 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     2161 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     2221 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     2281 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     2341 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     2401 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     2461 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     2521 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     2581 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     2641 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     2701 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     2761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2881 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     2941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
     3001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     3061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3661 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4261 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     4321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt
     4981 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5041 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5101 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5161 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5221 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5281 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5341 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5401 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cagtttaaac ttaagcttac
     5461 catggttcca gaccgtgaca atgatggaat ccctgattca ttagaggtag aaggatatac
     5521 ggttgatgtc aaaaataaaa gaacttttct ttcaccatgg atttctaata ttcatgaaaa
     5581 gaaaggatta accaaatata aatcatctcc tgaaaaatgg agcacggctt ctgatccgta
     5641 cagtgatttc gaaaaggtta caggacggat tgataagaat gtatcaccag aggcaagaca
     5701 cccccttgtg gcagcttatc cgattgtaca tgtagatatg gagaatatta ttctctcaaa
     5761 aaatgaggat caatccacac agaatactga tagtcaaacg agaacaataa gtaaaaatac
     5821 ttctacaagt aggacacata ctagtgaagt acatggaaat gcagaagtgc atgcgtcgtt
     5881 ctttgatatt ggtgggagtg tatctgcagg atttagtaat tcgaattcaa gtacggtcgc
     5941 aattgatcat tcactatctc tagcagggga aagaacttgg gctgaaacaa tgggtttaaa
     6001 taccgctgat acagcaagat taaatgccaa tattagatat gtaaatactg ggacggctcc
     6061 aatctacaac gtgttaccaa cgacttcgtt agtgttagga aaaaatcaaa cactcgcgac
     6121 aattaaagct aaggaaaacc aattaagtca aatacttgca cctaataatt attatccttc
     6181 taaaaacttg gcgccaatcg cattaaatgc acaagacgat ttcagttcta ctccaattac
     6241 aatgaattac aatcaatttc ttgagttaga aaaaacgaaa caattaagat tagatacgga
     6301 tcaagtatat gggaatatag caacatacaa ttttgaaaat ggaagagtga gggtggatac
     6361 aggctcgaac tggagtgaag tgttaccgca aattcaagaa acaactgcac gtatcatttt
     6421 taatggaaaa gatttaaatc tggtagaaag gcggatagcg gcggttaatc ctagtgatcc
     6481 attagaaacg actaaaccgg atatgacatt aaaagaagcc cttaaaatag catttggatt
     6541 taacgaatcg aatggaaact tacaatatca agggaaagac ataaccgaat ttgattttaa
     6601 tttcgatcaa caaacatctc aaaatatcaa gaatcagtta gcggaattaa acgtaactaa
     6661 catatatact gtattagata aaatcaaatt aaatgcaaaa atgaatattt taataagaga
     6721 taaacgtttt cattatgata gaaataacat agcagttggg gcggatgagt cagtagttaa
     6781 ggaggctcat agagaagtaa ttaattcgtc aacagaggga ttattgttaa atattgataa
     6841 ggatataaga aaaatattat caggttatat tgtagaaatt gaagatactg aagggcttaa
     6901 agaagttata aatgacagat atgatatgtt gaatatttct agtttacggc aagatggaaa
     6961 aacatttata gattttaaaa aatataatga taaattaccg ttatatataa gtaatcccaa
     7021 ttataaggta aatgtatatg ctgttactaa agaaaacact attattaatc ctagtgagaa
     7081 tggggatact agtaccaacg ggatcaagaa aattttaatc ttttctaaaa aaggctatga
     7141 gataggataa ggtaattgaa gctcgagtct agagggcccg tt
//
