LOCUS       sequence-198907-        4346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encoding two copies of T. thermophilus SecG for expression of SecYEG
            complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4346)
  AUTHORS   Sugano Y, Furukawa A, Nureki O, Tanaka Y, Tsukazaki T
  TITLE     SecY-SecA fusion protein retains the ability to mediate protein 
            transport.
  JOURNAL   PLoS One. 2017 Aug 18;12(8):e0183434. doi: 
            10.1371/journal.pone.0183434. eCollection 2017.
  PUBMED    28820900
REFERENCE   2  (bases 1 to 4346)
  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: "PLoS One.";
            date: "2017-08-18"; volume: "12(8):e0183434. doi"; pages: " 
            10.1371/journal.pone.0183434. eCollection 2017"
COMMENT     Sequence Label: pAK22
FEATURES             Location/Qualifiers
     source          1..4346
                     /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)."
     promoter        370..388
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    389..413
                     /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             702..746
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(784..802)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      798..845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1066..1725)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1640..1659
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1726..1828)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2220..2237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2354..2899)
                     /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(2434..2453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3124..4206)
                     /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     4168..4187
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4207..4284)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4330..4346,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggacctgc tctacacctt ggtcatcctc ttttacctgg gcgtggccgg
      121 gctattggtc tacctcgtgc tggtgcagga gcccaagcag ggggccgggg acctcatggg
      181 gggctcggcc gacctcttct ccgcccgggg cgtgacggga gggctctacc gcctcacggt
      241 gatcctgggg gtggtcttcg cggccctggc cttggtcatc ggcctctggc cccgttgaaa
      301 gcttgcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat
      361 aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tccccatctt
      421 agtatattag ttaagtataa gaaggagata tacatatgga cctgctctac accttggtca
      481 tcctctttta cctgggcgtg gccgggctat tggtctacct cgtgctggtg caggagccca
      541 agcagggggc cggggacctc atggggggct cggccgacct cttctccgcc cggggcgtga
      601 cgggagggct ctaccgcctc acggtgatcc tgggggtggt cttcgcggcc ctggccttgg
      661 tcatcggcct ctggccccgt tgaggtaccc tcgagtctgg taaagaaacc gctgctgcga
      721 aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg
      781 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      841 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
      901 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
      961 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
     1021 gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     1081 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     1141 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     1201 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     1261 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     1321 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     1381 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     1441 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     1501 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     1561 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     1621 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     1681 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     1741 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     1801 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     1861 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     1921 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
     1981 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
     2041 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
     2101 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     2161 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     2221 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     2281 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     2341 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     2401 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     2461 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     2521 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     2581 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     2641 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     2701 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     2761 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     2821 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     2881 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
     2941 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     3001 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     3061 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     3121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     3241 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     3301 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     3361 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     3421 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     3481 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     3541 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     3601 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     3661 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     3721 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     3781 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     3841 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     3901 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     3961 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     4021 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     4081 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     4141 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     4201 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     4261 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     4321 ttaggaaatt aatacgactc actata
//
