LOCUS       sequence-306050-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shows that expression of the protein of interest from Salmonella is 
            disrupted when SptP signal is removed from the pCASP-HilA vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  AUTHORS   Chabloz A, Schaefer JV, Kozieradzki I, Cronin SJF, Strebinger D, 
            Macaluso F, Wald J, Rabbitts TH, Pluckthun A, Marlovits TC, 
            Penninger JM
  TITLE     Salmonella-based platform for efficient delivery of functional 
            binding proteins to the cytosol.
  JOURNAL   Commun Biol. 2020 Jul 3;3(1):342. doi: 10.1038/s42003-020-1072-4.
  PUBMED    32620833
REFERENCE   2  (bases 1 to 6969)
  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: "Commun 
            Biol."; date: "2020-07-3"; volume: "3(1):342. doi"; pages: " 
            10.1038/s42003-020-1072-4"
COMMENT     Sequence Label: pCASP-3G124-HilA
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1037..1102
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1115..1186
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1212..1298
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1406..2284)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2311..2595
                     /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     2518..2537
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    4839..4897
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4930..4947)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5100..5171
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5328..5916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5408..5427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6004..6098)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6119..6778)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6693..6712
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6779..6881)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     6947..6965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgtcttcac ctcgagccac aagaaaacga ggtacggcat tgagccgcgt aaggcagtag
       61 cgatgtattc attgggcgtt ttttgaatgt tcactaacca ccgtcggggt ttaataactg
      121 catcagataa acgcagtcgt taagttctac aaagtcggtg acagataaca ggagtaagta
      181 atgcaagcac accaggatat tatcgctaat attggtgaga aattgggttt accgctcact
      241 tttgacgaca acaatcagtg cttattatta ctcgatagcg atatttttac gtctattgaa
      301 gctaaagatg atatctggtt attgaacggt atgattatac cgttatcgcc tgtttgtggc
      361 gattctatct ggcggcagat tatggtgatt aatggtgaac tggctgcgaa taatgaaggt
      421 acgttagcgt atattgatgc cgcagagacg ttgttgctta tacatgcaat taccgatctg
      481 acaaatactt accatattat atcgcagctt gagtcatttg tgaatcagca ggaagcgctc
      541 aaaaacatac tgcaggaatg acctgggcaa aaaattgctt gaagccgcac gcgcagggca
      601 ggatgacgag gtgcgcatcc ttatggcaaa tggggctgat gtcaatgccg cagatgatgt
      661 cggcgttact ccccttcatt tggctgcgca acgcggccac ttggagatcg tagaggtctt
      721 attaaagtat ggtgccgacg tgaatgctgc agacctttgg ggtcagacac cgttacattt
      781 ggcggccaca gcggggcact tggagatcgt ggaggtactg cttaagaacg gcgcggatgt
      841 caacgcacgg gataatatcg ggcacacccc cttgcacttg gctgcctggg caggccacct
      901 tgagattgta gaggtccttt tgaagtatgg cgctgacgtg aatgctcagg ataagtttgg
      961 caagacaccc tttgaccttg ccatcgataa tggtaacgag gatattgctg aagtcctgca
     1021 aaaggcagcc gtcgacgatt ataaagatca tgatggcgat tataaagatc atgatattga
     1081 ttataaagat gatgatgata aataataatc tagagattat aaagatgacg atgacaagga
     1141 ttataaagat gacgatgaca aggattataa agatgacgat gacaagtaat aaagcggccg
     1201 ctgattggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggacttt cgttttatct
     1261 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag accgatgcat
     1321 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     1381 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     1441 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     1501 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     1561 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     1621 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     1681 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     1741 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     1801 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     1861 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     1921 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     1981 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     2041 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     2101 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     2161 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     2221 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     2281 ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac
     2341 attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta
     2401 ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg
     2461 tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg
     2521 ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct
     2581 ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcggt acccccgagc
     2641 ccgtagaata tgacattaag ctcataataa aagctcaacc tgaccgttag tactaacagc
     2701 agaattactg aaacagtaga ttctatccta acgacttgta ttagttatta taacttttca
     2761 ccctgtaaga gaatacacta ttatcatgcc acattttaat cctgttcctg tatcgaataa
     2821 aaaattcgtc tttgatgatt tcatactcaa catggacggc tccctgctac gctcagaaaa
     2881 gaaagtcaat attccgccaa aagaatatgc cgttctggtc atcctgctcg aagccgccgg
     2941 cgagattgtg agtaaaaaca ccttactgga ccaggtatgg ggcgacgcgg aagttaacga
     3001 agaatctctt acccgctgta tttatgcctt acgacgtatt ctgtcggaag ataaagagca
     3061 tcgttacatt gaaacactgt acggacaggg ctatcggttt aatcgtccgg tcgtagtggt
     3121 gtctccgcca gcgccgcaac ctacgactca tacattggcg atacttcctt ttcagatgca
     3181 ggatcaggtt caatccgaga gtctgcatta ctctatcgtg aagggattat cgcagtatgc
     3241 gccctttggc ctgagcgtgc tgccggtgac cattacgaag aactgccgca gtgttaagga
     3301 tattcttgag ctcatggatc aattacgccc cgattattat atctccgggc agatgatacc
     3361 cgatggtaat gataatattg tacagattga gatagttcgg gttaaaggtt atcacctgct
     3421 gcaccaggaa agcattaagt tgatagaaca ccaacccgct tctctcttgc aaaacaaaat
     3481 tgcgaatctt ttgctcagat gtattcccgg acttcgctgg gacacaaagc agattagcga
     3541 gctaaattcg attgacagta ctatggttta cttacgcggt aagcatgagt taaatcaata
     3601 caccccctat agcttacagc aagcgcttaa attgctgact caatgcgtta acatgtcgcc
     3661 aaacagcatt gcgccttact gtgcgctggc agaatgctac ctcagcatgg cgcaaatggg
     3721 gatttttgat aaacaaaacg ctatgatcaa agctaaagaa catgcgatta aggcgacaga
     3781 gctggaccac aataatccac aagctttagg attactgggg ctaattaata cgattcactc
     3841 agaatacatc gtcgggagtt tgctattcaa acaagctaac ttactttcgc ccatttctgc
     3901 agatattaaa tattattatg gctggaatct tttcatggct ggtcagttgg aggaggcctt
     3961 acaaacgatt aacgagtgtt taaaattgga cccaacgcgc gcagccgcag ggatcactaa
     4021 gctgtggatt acctattatc ataccggtat tgatgatgct atacgtttag gcgatgaatt
     4081 acgctcacaa cacctgcagg ataatccaat attattaagt atgcaggtta tgtttctttc
     4141 gcttaaaggt aaacatgaac tggcacgaaa attaactaaa gaaatatcca cgcaggaaat
     4201 aacaggactt attgctgtta atcttcttta cgctgaatat tgtcagaata gtgagcgtgc
     4261 cttaccgacg ataagagaat ttctggaaag tgaacagcgt atagataata atccgggatt
     4321 attaccgtta gtgctggttg cccacggcga agctattgcc gagaaaatgt ggaataaatt
     4381 taaaaacgaa gacaatattt ggttcaaaag atggaaacag gatccccgct tgattaaatt
     4441 acggtaaaat ctgagagagg agatatgcat tattttttta tcatcgtaat ctggttgctt
     4501 agcataaata cggcatgggc tgattgctgg cttcaggctg aaaaaatgtt caatattgaa
     4561 tccgaactac tttacgctat cgcccagcag gaatcggcga tgaaacctgg cgccattggt
     4621 cataaccgag atggttcaac cgatcttggc ctgatgcaaa ttaacagctt ccatatgaaa
     4681 aggctgaaaa aaatggggat tagtgaaaaa cagttgttac aggacccctg catttctgtc
     4741 attgtgggcg cctccatttt atcagatatg atgaaaatct acggttatag ctgggaggcc
     4801 gttggcgctt ataatgccgg gacgtcgccg aaacgatcga attcgagctc ggtacccggg
     4861 gggatcctct agagtcgacc tgcaggcatg caagcttggc tgttttggcg gatgagagaa
     4921 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     4981 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     5041 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     5101 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     5161 tgaacgctct cctgagtcta gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5761 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gactagtgct
     6001 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     6061 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     6121 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     6181 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     6241 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     6301 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     6361 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     6421 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     6481 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     6541 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     6601 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     6661 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     6721 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     6781 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     6841 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaga
     6901 tatcgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6961 gaggccctt
//
