LOCUS       sequence-206084-        3845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express periplasmic, outer membrane, or secreted proteins in E. coli
            - OmpA signal and His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3845)
  AUTHORS   Casasanta MA, Slade DJ
  TITLE     A Vector Suite for the Overexpression and Purification of Tagged 
            Outer Membrane, Periplasmic, and Secreted Proteins in E. coli.
  JOURNAL   Methods Mol Biol. 2019;1960:123-138. doi: 
            10.1007/978-1-4939-9167-9_11.
  PUBMED    30798527
REFERENCE   2  (bases 1 to 3845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2019"; volume: "1960:123-138. doi"; pages: " 
            10.1007/978-1-4939-9167-9_11"
COMMENT     Sequence Label: pDJSVT87
FEATURES             Location/Qualifiers
     source          1..3845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     sig_peptide     77..139
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             158..175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             203..247
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(285..303)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      299..346
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(566..1225)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1140..1159
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1226..1328)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1720..1737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1854..2398)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1934..1953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2623..3705)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3667..3686
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3706..3783)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3829..3845,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3829..3845,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggaatga aaaagacagc tatcgcgatt gcagtggcac tggctggttt
      121 cgctaccgta gcgcaggccg ctccgaaaga taacacccac caccaccacc accacgcggc
      181 cgcgggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
      241 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
      301 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
      361 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
      421 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
      481 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcacca ggcgtttaag
      541 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
      601 gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga tgaacctgaa
      661 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccata gtgaaaacgg
      721 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
      781 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
      841 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
      901 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
      961 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag
     1021 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     1081 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     1141 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     1201 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     1261 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     1321 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     1381 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
     1441 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
     1501 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
     1561 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
     1621 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
     1681 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
     1741 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     1801 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     1861 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     1921 ccgacaggac tataaagata ccaggcgttt cccctggcgg ctccctcgtg cgctctcctg
     1981 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     2041 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     2101 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     2161 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     2221 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     2281 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     2341 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     2401 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa
     2461 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     2521 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     2581 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     2641 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2701 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2761 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2821 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2881 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     2941 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     3001 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     3061 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     3121 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     3181 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     3241 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     3301 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     3361 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     3421 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     3481 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     3541 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     3601 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     3661 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3721 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     3781 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     3841 ctata
//
