LOCUS       sequence-215577-        3844 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 3844)
  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 3844)
  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..3844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    46..70
                     /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             198..215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             243..287
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(325..343)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      339..386
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(607..1266)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1181..1200
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1267..1369)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1761..1778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1895..2440)
                     /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(1975..1994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2665..3747)
                     /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     3709..3728
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3748..3825)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat accatgggac
      121 gctatatccg acagttatgc tgtgtaagct tactctgctt aagcggatct gccgtcgccg
      181 cgaacgtccg tctacagcac caccaccacc accacgcggc cgcgggtacc ctcgagtctg
      241 gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag
      301 cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      361 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
      421 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
      481 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
      541 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
      601 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
      661 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
      721 cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
      781 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
      841 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
      901 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
      961 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1021 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
     1081 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1141 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1201 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1261 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1321 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     1381 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     1441 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     1501 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     1561 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     1621 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     1681 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     1741 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     1801 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     1861 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     1921 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     1981 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     2041 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     2101 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     2161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     2221 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     2281 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     2341 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     2401 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     2461 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     2521 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     2581 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     2641 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2701 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     2761 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     2821 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     2881 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     2941 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     3001 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     3061 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     3121 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     3181 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     3241 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     3301 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     3361 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     3421 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     3481 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     3541 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     3601 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     3661 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     3721 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     3781 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     3841 ctcc
//
