LOCUS       sequence-346121-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP preceeded by the signal peptide of TorA (spTorA) with an 
            N-terminal 6xHis tag and TEV cleavage site in pBAD24..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Bageshwar UK, DattaGupta A, Musser SM
  TITLE     Influence of the TorD signal peptide chaperone on Tat-dependent 
            protein translocation.
  JOURNAL   PLoS One. 2021 Sep 9;16(9):e0256715. doi: 
            10.1371/journal.pone.0256715. eCollection 2021.
  PUBMED    34499687
REFERENCE   2  (bases 1 to 5412)
  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: "2021-09-9"; volume: "16(9):e0256715. doi"; pages: " 
            10.1371/journal.pone.0256715. eCollection 2021"
COMMENT     Sequence Label: pH6-TEV-spTorA-GFP(C)
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1326..1343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1344..1364
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1500..2207
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNHHNVYITADKQKNGIKANF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLDTHSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     CDS             1500..2207
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNHHNVYITADKQKNGIKANF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLDTHSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(1524..1552)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2147..2167
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2270..2287)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2440..2526
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2618..2645
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2664..2755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2756..2824,2825..3616)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2974..2993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3658..4113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3745..3764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3955..3976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4224..4812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4713..4732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4966..4983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5149..5171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5309..5328
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tggcgcatca tcatcatcat catgaaaatc tctactttca aggcggcgcg aacaataacg
     1381 atctctttca ggcatcacgt cggcgttttc tggcacaact cggcggctta accgtcgccg
     1441 ggatgctggg gccgtcattg ttaacgccgc gacgtgcgac tgcggcgcaa gcggcgcgta
     1501 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     1561 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     1621 cccttaaatt tatttgcact actggaaaac tacctgttcc ctggccaaca cttgtcacta
     1681 ctttcggtta tggtgttcaa tgctttgcga gatacccaga tcatatgaaa cggcatgact
     1741 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     1801 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     1861 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttgggcac aaattggaat
     1921 acaactataa ccaccacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
     1981 ctaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     2041 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctggaca
     2101 cacactctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     2161 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaactc gagtgctaac
     2221 tgcaggcatg caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     2281 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     2341 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     2401 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     2461 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2521 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     2581 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     2641 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     2701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg
     3661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3841 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3961 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4081 cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt
     4141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4981 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5041 caccgcatag ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     5101 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5161 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagaaggag atggcgccca
     5221 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5281 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     5341 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt
     5401 tgacagctta tc
//
