LOCUS       40924_5744        4808 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBAD-HisB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4808)
  AUTHORS   Zhang W, Lohman AW, Zhuravlova Y, Lu X, Wiens MD, Hoi H, Yaganoglu 
            S, Mohr MA, Kitova EN, Klassen JS, Pantazis P, Thompson RJ, Campbell
            RE.
  TITLE     Optogenetic control with a photocleavable protein, PhoCl
  JOURNAL   Nat. Methods (2017) In press
  PUBMED    28288123
REFERENCE   2  (bases 1 to 4808)
  AUTHORS   Zhang W.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-FEB-2017) Chemistry, University of Alberta, 11227 
            Saskatchewan Drive, Department of Chemistry, CCIS 4-140, Edmonton, 
            Alberta T6G2C5, Canada
REFERENCE   3  (bases 1 to 4808)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4808)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-FEB-2017) Chemistry, University of Alberta, 11227 Saskatchewan 
            Drive, Department of Chemistry, CCIS 4-140, Edmonton, Alberta 
            T6G2C5, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: GeneStudio v. 2.2.0.0
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             331..348
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             388..411
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             427..1152
                     /codon_start=1
                     /label=PhoCl
                     /note="photocleavable protein that separates into two
                     fragments upon exposure to violet light (Zhang et al., 
                     2017)"
                     /translation="VIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKI
                     HFKGTNFPPNGPVMQKRTVGWEASTEKMYERDGVLKGDVKMKLLLKGGGHYRCDYRTTY
                     KVKQKPVKLPDYHFVDHRIEILSHDKDYNKVKLYEHAVARNSTDSMDELYKGGSGGMVS
                     KGEETITSVIKPDMKNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFA
                     YDILTTAFHYGNRVFTKYPR"
     terminator      1379..1465
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1557..1584
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1603..1694
                     /label=AmpR promoter
     CDS             1695..2552
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2726..3314
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3500..3640)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3907..4782)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagtgtga tccctgacta cttcaagcag agcttccccg agggctacag ctgggagcgc
      481 agcatgacct acgaggacgg cggcatctgc atcgccacca acgacatcac aatggagggg
      541 gacagcttca tcaacaagat ccacttcaag ggcacgaact tcccccccaa cggccccgtg
      601 atgcagaaga ggaccgtggg ctgggaggcc agcaccgaga agatgtacga gcgcgacggc
      661 gtgctgaagg gcgacgtgaa gatgaagctg ctgctgaagg gcggcggcca ctatcgctgc
      721 gactaccgca ccacctacaa ggtcaagcag aagcccgtaa agctgcccga ctaccacttc
      781 gtggaccacc gcatcgagat cctgagccac gacaaggact acaacaaggt gaagctgtac
      841 gagcacgccg tggcccgcaa ctccaccgac agcatggacg agctgtacaa gggtggcagc
      901 ggtggcatgg tgagcaaggg cgaggagacc attacaagcg tgatcaagcc tgacatgaag
      961 aacaagctgc gcatggaggg caacgtgaac ggccacgcct tcgtgatcga gggcgagggc
     1021 agcggcaagc ccttcgaggg catccagacg attgatttgg aggtgaagga gggcgccccg
     1081 ctgcccttcg cctacgacat cctgaccacc gccttccact acggcaaccg cgtgttcacc
     1141 aagtacccac ggggaggtgg aggtacctaa aagcttggct gttttggcgg atgagagaag
     1201 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     1261 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     1321 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     1381 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1441 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     1501 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     1561 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     1621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1681 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1741 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1801 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1861 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1921 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1981 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2041 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2101 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2161 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2221 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2281 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2341 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2401 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2461 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2521 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2581 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2641 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3361 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3421 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3481 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3541 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3601 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3661 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3721 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3781 gcagatcaat tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga
     3841 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
     3901 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     3961 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     4021 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     4081 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     4141 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     4201 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     4261 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     4321 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     4381 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     4441 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     4501 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     4561 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     4621 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     4681 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     4741 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     4801 attcagag
//
