LOCUS       Exported                4701 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  ORF selection and expression vector pSOS-X, complete sequence.
ACCESSION   EF030426
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4701)
  AUTHORS   Cox JC, Lape J, Sayed MA, Hellinga HW.
  TITLE     Protein fabrication automation
  JOURNAL   Protein Sci. 16 (3), 379-390 (2007)
  PUBMED    17242375
REFERENCE   2  (bases 1 to 4701)
  AUTHORS   Cox JC, Hellinga HW.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2006) Biochemistry, Duke University Medical 
            Center, Research Drive, DUMC 3711, Nanaline 413, Durham, NC 27710, 
            USA
REFERENCE   3  (bases 1 to 4701)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4701
                     /organism="ORF selection and expression vector pSOS-X"
                     /mol_type="other DNA"
                     /db_xref="taxon:409551"
     rep_origin      complement(240..828)
                     /direction=LEFT
                     /label=pMB1 plasmid replication origin
                     /note="pMB1 plasmid replication origin"
     gene            complement(999..1929)
                     /gene="bla"
                     /label=bla
     CDS             complement(999..1859)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="ABK62682.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1860..1964)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(1867..1871)
                     /regulatory_class="ribosome_binding_site"
                     /gene="bla"
                     /note="Shine-Delgarno sequence"
     regulatory      complement(1903..1929)
                     /regulatory_class="promoter"
                     /gene="bla"
     gene            2106..3254
                     /gene="lacI"
                     /label=lacI
     regulatory      2106..2151
                     /regulatory_class="promoter"
                     /gene="lacI"
     CDS             2172..3254
                     /codon_start=1
                     /gene="lacI"
                     /product="lactose operon repressor"
                     /label=lacI
                     /note="DNA binding protein"
                     /protein_id="ABK62683.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        3503..3531
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      3503..3530
                     /regulatory_class="promoter"
                     /note="Ptac promoter"
     protein_bind    3539..3555
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      3611..3616
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Delgarno sequence"
     misc_feature    3617..3624
                     /label=PacI restriction site
                     /note="PacI restriction site"
     misc_feature    3633..3638
                     /label=NaeI restriction site
                     /note="NaeI restriction site"
     misc_feature    3639..3656
                     /label=hexahistidine protein purification tag
                     /note="hexahistidine protein purification tag"
     CDS             3639..3656
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3657..3659
                     /label=amber stop codon
                     /note="amber stop codon"
     CDS             3666..4322
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /EC_number="2.3.1.28"
                     /label=cat
                     /note="chloramphenicol resistance; fused to the 3 end of 
                     the inserted gene; inserted gene provides canonical 
                     methionine start codon"
                     /protein_id="ABK62684.1"
                     /translation="EKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLK
                     TVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSL
                     WSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMD
                     NFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            3666..4322
                     /gene="cat"
                     /label=cat
     terminator      4492..4578
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      4495..4698
                     /regulatory_class="terminator"
                     /note="rrnB T1T2 transcriptional terminator"
     terminator      4670..4697
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"