LOCUS       sequence-297915-        6964 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 6964)
  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 6964)
  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..6964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1027..1092
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1105..1176
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1202..1288
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1396..2274)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2301..2585
                     /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     2508..2527
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    4824..4882
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4915..4932)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5085..5156
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5313..5901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5393..5412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5989..6083)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6104..6763)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6678..6697
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6764..6866)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     6932..6950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagccac aagaaaacga ggtacggcat tgagccgcgt aaggcagtag cgatgtattc
       61 attgggcgtt ttttgaatgt tcactaacca ccgtcggggt ttaataactg catcagataa
      121 acgcagtcgt taagttctac aaagtcggtg acagataaca ggagtaagta atgcaagcac
      181 accaggatat tatcgctaat attggtgaga aattgggttt accgctcact tttgacgaca
      241 acaatcagtg cttattatta ctcgatagcg atatttttac gtctattgaa gctaaagatg
      301 atatctggtt attgaacggt atgattatac cgttatcgcc tgtttgtggc gattctatct
      361 ggcggcagat tatggtgatt aatggtgaac tggctgcgaa taatgaaggt acgttagcgt
      421 atattgatgc cgcagagacg ttgttgctta tacatgcaat taccgatctg acaaatactt
      481 accatattat atcgcagctt gagtcatttg tgaatcagca ggaagcgctc aaaaacatac
      541 tgcaggaatg acctgggcaa aaaattgctt gaagccgcac gcgcagggca ggatgacgag
      601 gtgcgcatcc ttatggcaaa tggggctgat gtcaatgccg cagatgatgt cggcgttact
      661 ccccttcatt tggctgcgca acgcggccac ttggagatcg tagaggtctt attaaagtat
      721 ggtgccgacg tgaatgctgc agacctttgg ggtcagacac cgttacattt ggcggccaca
      781 gcggggcact tggagatcgt ggaggtactg cttaagaacg gcgcggatgt caacgcacgg
      841 gataatatcg ggcacacccc cttgcacttg gctgcctggg caggccacct tgagattgta
      901 gaggtccttt tgaagtatgg cgctgacgtg aatgctcagg ataagtttgg caagacaccc
      961 tttgaccttg ccatcgataa tggtaacgag gatattgctg aagtcctgca aaaggcagcc
     1021 gtcgacgatt ataaagatca tgatggcgat tataaagatc atgatattga ttataaagat
     1081 gatgatgata aataataatc tagagattat aaagatgacg atgacaagga ttataaagat
     1141 gacgatgaca aggattataa agatgacgat gacaagtaat aaagcggccg ctgattggca
     1201 tcaaataaaa cgaaaggctc agtcgaaaga ctgggacttt cgttttatct gttgtttgtc
     1261 ggtgaacgct ctcctgagta ggacaaatcc gccgccctag accgatgcat aatgtgcctg
     1321 tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt
     1381 ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     1441 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     1501 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     1561 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     1621 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     1681 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     1741 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     1801 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     1861 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     1921 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     1981 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     2041 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     2101 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     2161 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     2221 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     2281 catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca
     2341 ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat
     2401 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
     2461 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
     2521 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
     2581 tccatacccg tttttttggg ctagcgaatt cgagctcggt acccgagccc gtagaatatg
     2641 acattaagct cataataaaa gctcaacctg accgttagta ctaacagcag aattactgaa
     2701 acagtagatt ctatcctaac gacttgtatt agttattata acttttcacc ctgtaagaga
     2761 atacactatt atcatgccac attttaatcc tgttcctgta tcgaataaaa aattcgtctt
     2821 tgatgatttc atactcaaca tggacggctc cctgctacgc tcagaaaaga aagtcaatat
     2881 tccgccaaaa gaatatgccg ttctggtcat cctgctcgaa gccgccggcg agattgtgag
     2941 taaaaacacc ttactggacc aggtatgggg cgacgcggaa gttaacgaag aatctcttac
     3001 ccgctgtatt tatgccttac gacgtattct gtcggaagat aaagagcatc gttacattga
     3061 aacactgtac ggacagggct atcggtttaa tcgtccggtc gtagtggtgt ctccgccagc
     3121 gccgcaacct acgactcata cattggcgat acttcctttt cagatgcagg atcaggttca
     3181 atccgagagt ctgcattact ctatcgtgaa gggattatcg cagtatgcgc cctttggcct
     3241 gagcgtgctg ccggtgacca ttacgaagaa ctgccgcagt gttaaggata ttcttgagct
     3301 catggatcaa ttacgccccg attattatat ctccgggcag atgatacccg atggtaatga
     3361 taatattgta cagattgaga tagttcgggt taaaggttat cacctgctgc accaggaaag
     3421 cattaagttg atagaacacc aacccgcttc tctcttgcaa aacaaaattg cgaatctttt
     3481 gctcagatgt attcccggac ttcgctggga cacaaagcag attagcgagc taaattcgat
     3541 tgacagtact atggtttact tacgcggtaa gcatgagtta aatcaataca ccccctatag
     3601 cttacagcaa gcgcttaaat tgctgactca atgcgttaac atgtcgccaa acagcattgc
     3661 gccttactgt gcgctggcag aatgctacct cagcatggcg caaatgggga tttttgataa
     3721 acaaaacgct atgatcaaag ctaaagaaca tgcgattaag gcgacagagc tggaccacaa
     3781 taatccacaa gctttaggat tactggggct aattaatacg attcactcag aatacatcgt
     3841 cgggagtttg ctattcaaac aagctaactt actttcgccc atttctgcag atattaaata
     3901 ttattatggc tggaatcttt tcatggctgg tcagttggag gaggccttac aaacgattaa
     3961 cgagtgttta aaattggacc caacgcgcgc agccgcaggg atcactaagc tgtggattac
     4021 ctattatcat accggtattg atgatgctat acgtttaggc gatgaattac gctcacaaca
     4081 cctgcaggat aatccaatat tattaagtat gcaggttatg tttctttcgc ttaaaggtaa
     4141 acatgaactg gcacgaaaat taactaaaga aatatccacg caggaaataa caggacttat
     4201 tgctgttaat cttctttacg ctgaatattg tcagaatagt gagcgtgcct taccgacgat
     4261 aagagaattt ctggaaagtg aacagcgtat agataataat ccgggattat taccgttagt
     4321 gctggttgcc cacggcgaag ctattgccga gaaaatgtgg aataaattta aaaacgaaga
     4381 caatatttgg ttcaaaagat ggaaacagga tccccgcttg attaaattac ggaatctgag
     4441 agaggagata tgcattattt ttttatcatc gtaatctggt tgcttagcat aaatacggca
     4501 tgggctgatt gctggcttca ggctgaaaaa atgttcaata ttgaatccga actactttac
     4561 gctatcgccc agcaggaatc ggcgatgaaa cctggcgcca ttggtcataa ccgagatggt
     4621 tcaaccgatc ttggcctgat gcaaattaac agcttccata tgaaaaggct gaaaaaaatg
     4681 gggattagtg aaaaacagtt gttacaggac ccctgcattt ctgtcattgt gggcgcctcc
     4741 attttatcag atatgatgaa aatctacggt tatagctggg aggccgttgg cgcttataat
     4801 gccgggacgt cgccgaaacg atcgaattcg agctcggtac ccggggggat cctctagagt
     4861 cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     4921 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     4981 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     5041 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     5101 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     5161 gtctagggcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5221 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5941 tggaacgaaa actcacgtta agggattttg gtcatgacta gtgcttggat tctcaccaat
     6001 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     6061 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     6121 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     6181 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     6241 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     6301 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     6361 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     6421 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     6481 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     6541 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     6601 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     6661 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     6721 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     6781 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     6841 agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccagatatcg acgtctaaga
     6901 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6961 tcac
//
