LOCUS       Exported                4208 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  ORF selection vector pSOS plasmid pSOS, complete sequence.
ACCESSION   EF030424
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4208)
  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 4208)
  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 4208)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4208
                     /organism="ORF selection vector pSOS"
                     /mol_type="other DNA"
                     /db_xref="taxon:409549"
     regulatory      70..75
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Delgarno sequence"
     misc_feature    76..83
                     /label=PacI restriction site
                     /note="PacI restriction site"
     misc_feature    92..97
                     /label=HpaI restriction site
                     /note="HpaI restriction site"
     CDS             116..772
                     /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="ABK62679.1"
                     /translation="EKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLK
                     TVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSL
                     WSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMD
                     NFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            116..772
                     /gene="cat"
                     /label=cat
     promoter        complement(846..864)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(871..887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1025..1327
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1538..2470
                     /gene="aphA1"
                     /label=aphA1
     regulatory      1538..1675
                     /regulatory_class="promoter"
                     /gene="aphA1"
     CDS             1676..2470
                     /codon_start=1
                     /gene="aphA1"
                     /product="aminoglycoside 3-phosphotransferase"
                     /EC_number="2.7.1.95"
                     /label=aphA1
                     /note="kanamycin resistance"
                     /protein_id="ABK62678.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      2671..2680
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2677..3051
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3163..3836
                     /label=pMB1 plasmid replication origin
                     /note="pMB1 plasmid replication origin"
     rep_origin      3189..3777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4053..4174
                     /regulatory_class="promoter"
                     /note="lactose operon promoter"
     protein_bind    4065..4086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4101..4131
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4139..4155
                     /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)."
     primer_bind     4163..4179
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4197..7
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"