LOCUS       sequence-297913-        7318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Triggers secretion of SptP120 fused to the E3_5 DARPin from 
            Salmonella upon induction with arabinose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7318)
  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 7318)
  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-SptP120-E3_5-HilA
FEATURES             Location/Qualifiers
     source          1..7318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1381..1446
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1459..1530
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1556..1642
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1750..2628)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2655..2939
                     /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     2862..2881
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    5178..5236
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5269..5286)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5439..5510
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6343..6437)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6458..7117)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7032..7051
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7118..7220)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7286..7304
                     /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 tgcaggaata tgctaaagta tgaggagaga aaattgaata atttaacgtt gtcttcgttt
      601 tcaaaagttg gtgtgtcgaa tgatgcccga ctttatattg ctaaggaaaa tactgataag
      661 gcatatgttg cgcctgaaaa attttcgtca aaagtattaa cctggcttgg aaaaatgccg
      721 ttatttaaaa acactgaagt ggtgcaaaaa catacggaaa atatcagagt acaggaccaa
      781 aagattttac agacatttct ccatgcacta acggaaaaat atggggaaac agcggttaat
      841 gacgcactgt taatgtcccg tataaatatg aacaaacccc ttacccaacg tttagcagtg
      901 cagatcacgg acctgggtaa gaaactgctg gaagctgctc gtgctggtca ggacgacgaa
      961 gttcgtatcc tgatggctaa cggtgctgac gttaacgcta ctgacaatga tggttatact
     1021 ccgctgcacc tggctgcttc taatggtcac ctggaaatcg ttgaagttct gctgaagaac
     1081 ggtgctgacg ttaacgcttc tgaccttact ggtattactc cgctgcacct ggctgctgct
     1141 actggtcacc tggaaatcgt tgaagttctg ctgaagcacg gtgctgacgt taacgcttat
     1201 gacaatgatg gtcatactcc gctgcacctg gctgctaagt atggtcacct ggaaatcgtt
     1261 gaagttctgc tgaagcacgg tgctgacgtt aacgctcagg acaaattcgg taagaccgct
     1321 ttcgacatct ccatcgacaa cggtaacgag gacctggctg aaatcctgca aaaggtcgac
     1381 gattataaag atcatgatgg cgattataaa gatcatgata ttgattataa agatgatgat
     1441 gataaataat aatctagaga ttataaagat gacgatgaca aggattataa agatgacgat
     1501 gacaaggatt ataaagatga cgatgacaag taataaagcg gccgctgatt ggcatcaaat
     1561 aaaacgaaag gctcagtcga aagactggga ctttcgtttt atctgttgtt tgtcggtgaa
     1621 cgctctcctg agtaggacaa atccgccgcc ctagaccgat gcataatgtg cctgtcaaat
     1681 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     1741 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     1801 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     1861 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     1921 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     1981 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     2041 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     2101 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     2161 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     2221 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     2281 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     2341 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     2401 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     2461 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     2521 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     2581 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     2641 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
     2701 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
     2761 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
     2821 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
     2881 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
     2941 cccgtttttt tgggctagcg aattcgagct cggtacccga gcccgtagaa tatgacatta
     3001 agctcataat aaaagctcaa cctgaccgtt agtactaaca gcagaattac tgaaacagta
     3061 gattctatcc taacgacttg tattagttat tataactttt caccctgtaa gagaatacac
     3121 tattatcatg ccacatttta atcctgttcc tgtatcgaat aaaaaattcg tctttgatga
     3181 tttcatactc aacatggacg gctccctgct acgctcagaa aagaaagtca atattccgcc
     3241 aaaagaatat gccgttctgg tcatcctgct cgaagccgcc ggcgagattg tgagtaaaaa
     3301 caccttactg gaccaggtat ggggcgacgc ggaagttaac gaagaatctc ttacccgctg
     3361 tatttatgcc ttacgacgta ttctgtcgga agataaagag catcgttaca ttgaaacact
     3421 gtacggacag ggctatcggt ttaatcgtcc ggtcgtagtg gtgtctccgc cagcgccgca
     3481 acctacgact catacattgg cgatacttcc ttttcagatg caggatcagg ttcaatccga
     3541 gagtctgcat tactctatcg tgaagggatt atcgcagtat gcgccctttg gcctgagcgt
     3601 gctgccggtg accattacga agaactgccg cagtgttaag gatattcttg agctcatgga
     3661 tcaattacgc cccgattatt atatctccgg gcagatgata cccgatggta atgataatat
     3721 tgtacagatt gagatagttc gggttaaagg ttatcacctg ctgcaccagg aaagcattaa
     3781 gttgatagaa caccaacccg cttctctctt gcaaaacaaa attgcgaatc ttttgctcag
     3841 atgtattccc ggacttcgct gggacacaaa gcagattagc gagctaaatt cgattgacag
     3901 tactatggtt tacttacgcg gtaagcatga gttaaatcaa tacaccccct atagcttaca
     3961 gcaagcgctt aaattgctga ctcaatgcgt taacatgtcg ccaaacagca ttgcgcctta
     4021 ctgtgcgctg gcagaatgct acctcagcat ggcgcaaatg gggatttttg ataaacaaaa
     4081 cgctatgatc aaagctaaag aacatgcgat taaggcgaca gagctggacc acaataatcc
     4141 acaagcttta ggattactgg ggctaattaa tacgattcac tcagaataca tcgtcgggag
     4201 tttgctattc aaacaagcta acttactttc gcccatttct gcagatatta aatattatta
     4261 tggctggaat cttttcatgg ctggtcagtt ggaggaggcc ttacaaacga ttaacgagtg
     4321 tttaaaattg gacccaacgc gcgcagccgc agggatcact aagctgtgga ttacctatta
     4381 tcataccggt attgatgatg ctatacgttt aggcgatgaa ttacgctcac aacacctgca
     4441 ggataatcca atattattaa gtatgcaggt tatgtttctt tcgcttaaag gtaaacatga
     4501 actggcacga aaattaacta aagaaatatc cacgcaggaa ataacaggac ttattgctgt
     4561 taatcttctt tacgctgaat attgtcagaa tagtgagcgt gccttaccga cgataagaga
     4621 atttctggaa agtgaacagc gtatagataa taatccggga ttattaccgt tagtgctggt
     4681 tgcccacggc gaagctattg ccgagaaaat gtggaataaa tttaaaaacg aagacaatat
     4741 ttggttcaaa agatggaaac aggatccccg cttgattaaa ttacggaatc tgagagagga
     4801 gatatgcatt atttttttat catcgtaatc tggttgctta gcataaatac ggcatgggct
     4861 gattgctggc ttcaggctga aaaaatgttc aatattgaat ccgaactact ttacgctatc
     4921 gcccagcagg aatcggcgat gaaacctggc gccattggtc ataaccgaga tggttcaacc
     4981 gatcttggcc tgatgcaaat taacagcttc catatgaaaa ggctgaaaaa aatggggatt
     5041 agtgaaaaac agttgttaca ggacccctgc atttctgtca ttgtgggcgc ctccatttta
     5101 tcagatatga tgaaaatcta cggttatagc tgggaggccg ttggcgctta taatgccggg
     5161 acgtcgccga aacgatcgaa ttcgagctcg gtacccgggg ggatcctcta gagtcgacct
     5221 gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     5281 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     5341 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     5401 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     5461 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtctag
     5521 ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg actagtgctt ggattctcac caataaaaaa
     6361 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     6421 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     6481 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     6541 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     6601 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     6661 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     6721 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     6781 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     6841 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     6901 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     6961 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     7021 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     7081 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     7141 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     7201 aacctcttac gtgccgatca acgtctcatt ttcgccagat atcgacgtct aagaaaccat
     7261 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
