LOCUS       sequence-211140-        3847 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 3847)
  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 3847)
  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..3847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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     94..156
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             175..192
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             220..264
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(302..320)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      316..363
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(584..1243)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1158..1177
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1244..1346)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1738..1755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1872..2417)
                     /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(1952..1971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2642..3724)
                     /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     3686..3705
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3725..3802)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggaatgaaaa agacagctat cgcgattgca
      121 gtggcactgg ctggtttcgc taccgtagcg caggccgctc cgaaagataa cacccaccac
      181 caccaccacc acgcggccgc gggtaccctc gagtctggta aagaaaccgc tgctgcgaaa
      241 tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc
      301 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      361 ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa
      421 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc
      481 gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
      541 agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc
      601 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
      661 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
      721 tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
      781 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
      841 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
      901 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
      961 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     1021 tacggaactc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     1081 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     1141 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     1201 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     1261 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     1321 agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct
     1381 tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag
     1441 gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga
     1501 tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct
     1561 actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta
     1621 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
     1681 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
     1741 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     1801 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     1861 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     1921 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     1981 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     2041 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     2101 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2161 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2221 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2281 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2341 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     2401 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag
     2461 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     2521 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     2581 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     2641 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2701 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     2761 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     2821 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     2881 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     2941 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     3001 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     3061 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     3121 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     3181 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     3241 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     3301 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     3361 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     3421 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     3481 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     3541 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     3601 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     3661 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     3721 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     3781 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     3841 aggaaat
//
