LOCUS       sequence-346120-        5385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP with the signal peptide of TorA (spTorA) in pBAD24. Includes a 
            C-terminal HHHHHHC (6xHisC) extension..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5385)
  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 5385)
  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: p-spTorA-GFP-H6C
FEATURES             Location/Qualifiers
     source          1..5385
                     /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             1455..2162
                     /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             1455..2162
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNHHNVYITADKQKNGIKANF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLDTHSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(1479..1507)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2102..2122
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2169..2186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2243..2260)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2413..2499
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2591..2618
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2637..2728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2729..2797,2798..3589)
                     /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(2947..2966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3631..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3928..3949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4197..4785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4686..4705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4939..4956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5122..5144
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5282..5301
                     /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 tggcgaacaa taacgatctc tttcaggcat cacgtcggcg ttttctggca caactcggcg
     1381 gcttaaccgt cgccgggatg ctggggccgt cattgttaac gccgcgacgt gcgactgcgg
     1441 cgcaagcggc gcgtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     1501 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     1561 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     1621 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
     1681 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     1741 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     1801 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     1861 ggcacaaatt ggaatacaac tataaccacc acaatgtata catcacggca gacaaacaaa
     1921 agaatggaat caaagctaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     1981 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     2041 accattacct ggacacacac tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     2101 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     2161 aactcgagca tcatcatcat catcattgct aactgcaggc atgcaagctt ggctgttttg
     2221 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     2281 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     2341 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     2401 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2461 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     2521 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     2581 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     2641 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2701 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2761 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2821 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2881 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2941 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     3001 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3061 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3121 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3181 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3241 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3301 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3361 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3421 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3481 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3541 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3601 agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg
     3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3841 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3961 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4081 caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4321 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4981 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtcatg gctgcgcccc
     5041 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5101 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5161 cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     5221 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     5281 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     5341 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatc
//
