LOCUS       sequence-206086-        3842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express periplasmic, outer membrane, or secreted proteins in E. coli
            - TamA signal and His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3842)
  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 3842)
  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: pDJSVT90
FEATURES             Location/Qualifiers
     source          1..3842
                     /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)."
     CDS             155..172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             200..244
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(282..300)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      296..343
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(563..1222)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1137..1156
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1223..1325)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1717..1734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1851..2395)
                     /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(1931..1950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2620..3702)
                     /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     3664..3683
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3703..3780)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3826..3842,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3826..3842,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggacgct atatccgaca gttatgctgt gtaagcttac tctgcttaag
      121 cggatctgcc gtcgccgcga acgtccgtct acagcaccac caccaccacc acgcggccgc
      181 gggtaccctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga
      241 ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc
      301 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
      361 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
      421 ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa
      481 tttctgccat tcatccgctt attatcactt attcaggcgt agcaccaggc gtttaagggc
      541 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
      601 attcattaag cattctgccg acatggaagc catcacagac ggcatgatga acctgaatcg
      661 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg aaaacggggg
      721 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
      781 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
      841 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
      901 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
      961 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     1021 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1081 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1141 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1201 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     1261 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     1321 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     1381 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     1441 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     1501 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     1561 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     1621 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     1681 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     1741 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     1801 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     1861 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     1921 acaggactat aaagatacca ggcgtttccc ctggcggctc cctcgtgcgc tctcctgttc
     1981 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     2041 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     2101 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     2161 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     2221 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     2281 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     2341 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     2401 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     2461 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     2521 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     2581 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2641 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2701 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     2761 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     2821 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     2881 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     2941 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3001 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3061 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3121 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     3181 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3241 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3301 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     3361 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     3421 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     3481 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     3541 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     3601 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     3661 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     3721 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     3781 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     3841 ta
//
