LOCUS       sequence-197570-        7761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encoding SecY(L2V, R252G)–(GGSG)4–SecA1-939–His10 fusion protein and
            SecE and SecG from Thermus thermophilus HB8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7761)
  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 7761)
  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: pTT610
FEATURES             Location/Qualifiers
     source          1..7761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..116
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    233..254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(269..274,275..292,293..299)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    307..323
                     /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)."
     primer_bind     312..334
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4523..4552
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(5016..5033)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5016..5032)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5025..5047)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5213..5231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5269..5291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5391..5410
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5427..5882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5564..5585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5776..5795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5982..6086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6087..6155,6156..6947)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6305..6324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7118..7706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7607..7626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
       61 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      121 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      181 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      241 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      301 gtggaattgt gagcggataa caatttcaca caggaaacag accatggtca aggccttctg
      361 gagcgccctg cagatccccg agctccgcca gcgggtgctc ttcaccctcc tcgtcctcgc
      421 cgcctaccgc ctcggggcct tcatccccac cccgggggtg gacctggaca agatccagga
      481 gttcctgcgc acggcccaag gcggggtctt cggcatcatc aacctcttct ccgggggcaa
      541 ctttgagcgc ttctccatct tcgccctggg catcatgccc tacatcaccg cggccatcat
      601 catgcagatc ctggtcacgg tggtgccggc gctggagaag ctctccaagg agggggagga
      661 ggggcggcgc atcatcaacc agtacacccg catcgggggc atcgccctgg gcgccttcca
      721 ggggttcttc ctggccacgg cctttttggg ggccgagggg ggccgcttcc tcctcccggg
      781 ctggtccccc ggccccttct tctggttcgt ggtggtggtg acccaggtgg cgggcatcgc
      841 cctcctcctg tggatggcgg agcgcatcac cgagtacggc atcggcaacg gcacgagcct
      901 catcatcttc gccgggatcg tggtggaatg gcttccccag atcctgcgca ccataggcct
      961 catccgcacc ggggaggtca acctggtggc cttcctcttc ttcctcgcct tcatcgtcct
     1021 ggccttcgcc ggcatggccg ccgtccagca ggccgagcgc aggatccccg tccagtacgc
     1081 ccgcaaggtg gtggggggca gggtctacgg gggccaggcc acctacatcc ccatcaagct
     1141 caacgccgcc ggggtgatcc ccatcatctt cgccgcggcc atcctgcaga tccccatctt
     1201 cctggccgcc cccttccagg acaaccccgt gctgcagggc atcgccaact tcttcaaccc
     1261 cacccggcct tcggggctct tcatagaggt cctcctcgtc atcctcttca cctacgtcta
     1321 caccgccgtc cagtttgacc ccaagcgcat cgccgagtcg ctgcgggagt acggggggtt
     1381 catcccgggg atccgcccgg gcgagcccac ggtgaagttc ctggagcata tcgtttcccg
     1441 tctcaccctc tggggggccc tcttccttgg gctcgtgacc ctccttcccc agatcatcca
     1501 gaacctcacc ggcatccaca gcatcgcctt ctcaggcatc ggcctcctca tcgtggtggg
     1561 cgtggccttg gacaccctga ggcaggtgga gagccagctc atgctcagga gctacgaggg
     1621 cttcctctcc cgcgggcgcc tgcgggggcg gaaccggggc ggtagcggcg gcggtagcgg
     1681 cggcggtagc ggcggcggta gcggcatgct aggtcttttg cgcaggctct ttgacaacaa
     1741 cgagcgggag atcgcccgct actacaagca ggtggtggag ccggtgaacc gcctcgaggc
     1801 cgaggtggag aagctccccg acctcgccgc cgcctaccgg gagctcaagg agaagcacga
     1861 gaagggggct tccctggacg agcttctccc catggccttc gccctcaccc gggagtccgc
     1921 caagcgctac ctgggcatgc gccactttga cgtccagctc atcggcgggg ccgtcctcca
     1981 cgagggcaag atcgccgaga tgaagacggg cgagggtaag accctggtgg ccaccctggc
     2041 cgtggccttg aacgccctca cgggcaaggg cgtccacgtg gtcacggtga acgactacct
     2101 ggcccgccgc gacgccgagt ggatggggcc cgtctaccgg ggcctgggcc tctccgtggg
     2161 ggtgatccag cacgcctcca ccccggcgga gcgccgcaag gcctacctcg ccgacgtcac
     2221 ctacgtgacc aactccgagc tcggctttga ctacctgcgg gacaacatgg ccataagccc
     2281 cgaccagctc gtcctccgcc acgaccaccc cctccactac gccatcattg acgaggtgga
     2341 ctccatcctc attgacgagg cccgcacccc cctcatcatc tctggccccg cggagaaggc
     2401 caccgacctc tactacaaga tggcggagat cgccaagaag ctggaaaggg gcctgcccgc
     2461 cgagcccggg gtgcgcaagg agcccacggg ggactacacc gttgaggaga agaaccgctc
     2521 cgtccacctc accctccagg gcatcgccaa ggcggagaag ctcctcggca ttgagggcct
     2581 cttcagtccg gagaacatgg agctcgccca catgctcatc caggccatcc gggcgaagga
     2641 gctctaccac cgggaccggg actacatcgt ccaggacggc caggtcatca tcgtggacga
     2701 gttcacgggc cgcctcatgc cgggccgccg ctacggggag ggcctccacc aggccattga
     2761 ggccaaggag ggggtccgga ttgagcggga gaaccagacc ctggccacca tcacctacca
     2821 gaacttcttc cgcctctacg agaagcgggc ggggatgacg ggcaccgcca agaccgagga
     2881 gaaggagttc caggagatct acgggatgga cgtggtggtg gtgcccacca accgcccggt
     2941 gatccgcaag gacttccccg acgtggtcta ccgcacggag aaggggaagt tctacgccgt
     3001 ggtggaggag atcgccgaga agtacgagcg gggccagccc gtcctcgtgg gcaccatcag
     3061 cattgagaag tcggaaaggc tctcccagat gctcaaggag cccaggctct acctccccag
     3121 gctggagatg cgcctggagc tcttcaagaa ggcgagccag aagcagcagg gcccggagtg
     3181 ggagaggctc aggaagcttt tggaaaggcc cgcccagctc aaggacgagg accttgcccc
     3241 ctttgagggg ctcatcccgc cgaaggggaa cctgaggacg gcctgggagg gcctgaagcg
     3301 ggcggtgcac accctcgccg tcctccgcca ggggatcccc caccaggtcc taaacgccaa
     3361 gcaccacgcc agggaagcgg agatcgtggc ccaggcgggg cggagcaaga cggtcaccat
     3421 cgccaccaac atggcggggc ggggcacgga catcaagctc gggggcaacc ccgagtacct
     3481 ggccgccgcc ctcctggaga aggaaggctt tgaccggtac gagtggaagg tggagctctt
     3541 catcaagaag atggtggcgg ggaaggagga ggaggccagg gccctcgccc aggagctcgg
     3601 gatccgggag gagcttctgg aaaggatccg cgagatccgg gaggagtcca agcaggacga
     3661 ggagcgggtc cgggccctgg gagggctttt catcatcggc accgagcggc acgagtcccg
     3721 ccgcatagac aaccagctcc gcggccgcgc cgggcgccag ggggaccccg ggggaagccg
     3781 cttctacgtc tcctttgacg acgacctcat gcgcctcttc gcctccgacc gggtcatcgc
     3841 catgctggac cggatgggct ttgacgactc cgagcccatt gagcacccca tggtgacccg
     3901 gtccattgag cgggcccaga agcgggtgga ggaccggaac ttcgccatcc gcaagcagct
     3961 tctccagttt gacgacgtct taagccgcca gcgggaggtc atctacgccc agcgccgcct
     4021 gattctcctc ggcaaggacg aggaggtgaa ggaggccgcc atcggcatgg tggaggagac
     4081 ggtggcttct ttggcggaga acttcctgaa ccccgaggtc caccccgagg actgggacct
     4141 cgagggcctc aaggccaccc tcctggacac cgcgccccag cttcaggact tcccctttgc
     4201 ggagcttagg gccttgaagg ccgaggaggc ggtggagagg ctcgtggagg ccgccctcaa
     4261 ggcctacgag gcccgcgagg ccgagctttc cccgcccctg atgcgggccg tggagcgctt
     4321 cgtcatcctc aacgtggtgg acaacgcctg gaaggagcac ctgcacaacc tggacgtcct
     4381 caggcagggg atcttcctgc ggggctacgg ccagaaggac cccttccagg agtacaagat
     4441 tgaggccacc cgcctcttca acgagatggt ggccttcatc aagagcgagg tggccaagtt
     4501 cctcttccgc ctcaaggtgg agcaccatca ccatcaccat caccatcacc attagtctag
     4561 aaggaggttt aaattatgtt cgcccggctg atccgttact tccaggaagc gcgggccgag
     4621 ctcgcccggg tcacctggcc cacgcgggag caggtcgtgg agggcacgca ggccatcctc
     4681 ctcttcacct tggccttcat ggtgatcctc gggctttacg acacggtctt ccgcttcctg
     4741 atagggcttc tgcgatagaa gcttaggagg tttaaattat ggacctgctc tacaccttgg
     4801 tcatcctctt ttacctgggc gtggccgggc tattggtcta cctcgtgctg gtgcaggagc
     4861 ccaagcaggg ggccggggac ctcatggggg gctcggccga cctcttctcc gcccggggcg
     4921 tgacgggagg gctctaccgc ctcacggtga tcctgggggt ggtcttcgcg gccctggcct
     4981 tggtcatcgg cctctggccc cgttgaaagc ttggcactgg ccgtcgtttt acaacgtcgt
     5041 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5101 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5161 aatggcgaat ggcgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5221 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5281 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5341 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5401 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     5461 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5521 ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5581 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5641 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     5701 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5761 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5821 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5881 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5941 taccgcatca ggcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6241 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7261 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7681 cgtcaggggg gcggagccca tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7741 ccttttgctg gccttttgct c
//
