LOCUS       sequence-306311-        7326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Triggers secretion of SptP120 fused to the K27 DARPin from 
            Salmonella upon induction with arabinose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  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 7326)
  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-K27-HilA
FEATURES             Location/Qualifiers
     source          1..7326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1383..1448
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1461..1532
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1558..1644
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1752..2630)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2657..2941
                     /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     2864..2883
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    5185..5243
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5276..5293)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5446..5517
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5674..6262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5754..5773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6350..6444)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6465..7124)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7039..7058
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7125..7227)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7293..7311
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgagccaca agaaaacgag gtacggcatt gagccgcgta aggcagtagc gatgtattca
       61 ttgggcgttt tttgaatgtt cactaaccac cgtcggggtt taataactgc atcagataaa
      121 cgcagtcgtt aagttctaca aagtcggtga cagataacag gagtaagtaa tgcaagcaca
      181 ccaggatatt atcgctaata ttggtgagaa attgggttta ccgctcactt ttgacgacaa
      241 caatcagtgc ttattattac tcgatagcga tatttttacg tctattgaag ctaaagatga
      301 tatctggtta ttgaacggta tgattatacc gttatcgcct gtttgtggcg attctatctg
      361 gcggcagatt atggtgatta atggtgaact ggctgcgaat aatgaaggta cgttagcgta
      421 tattgatgcc gcagagacgt tgttgcttat acatgcaatt accgatctga caaatactta
      481 ccatattata tcgcagcttg agtcatttgt gaatcagcag gaagcgctca aaaacatact
      541 gcaggaatat gctaaagtat gaggagagaa aattgaataa tttaacgttg tcttcgtttt
      601 caaaagttgg tgtgtcgaat gatgcccgac tttatattgc taaggaaaat actgataagg
      661 catatgttgc gcctgaaaaa ttttcgtcaa aagtattaac ctggcttgga aaaatgccgt
      721 tatttaaaaa cactgaagtg gtgcaaaaac atacggaaaa tatcagagta caggaccaaa
      781 agattttaca gacatttctc catgcactaa cggaaaaata tggggaaaca gcggttaatg
      841 acgcactgtt aatgtcccgt ataaatatga acaaacccct tacccaacgt ttagcagtgc
      901 agatcacgga tcttggaaaa aaattacttg aggcggcacg cgcagggcag gatgacgagg
      961 tacgtatcct tatggcgaat ggagccgatg tgaatgctca cgacactttc ggtttcactc
     1021 ctcttcacct tgctgccttg tacggacact tagagatcgt cgaggtatta ttaaagaatg
     1081 gggcagacgt gaatgcggat gactcatatg gtcgtactcc cctgcatctg gcagcgatgc
     1141 gtgggcacct tgagattgta gaagtactgc tgaaatacgg ggcagacgta aatgccgcag
     1201 atgaggaggg acgcacacct ttgcatcttg cagcaaaacg tggccacttg gaaattgttg
     1261 aagtactgct gaagaacggg gcagatgtta acgcacagga taaattcggc aaaacagctt
     1321 tcgatatttc cattgataat ggcaatgagg atcttgcgga aattctgcag aagcttgtcg
     1381 acgattataa agatcatgat ggcgattata aagatcatga tattgattat aaagatgatg
     1441 atgataaata ataatctaga gattataaag atgacgatga caaggattat aaagatgacg
     1501 atgacaagga ttataaagat gacgatgaca agtaataaag cggccgctga ttggcatcaa
     1561 ataaaacgaa aggctcagtc gaaagactgg gactttcgtt ttatctgttg tttgtcggtg
     1621 aacgctctcc tgagtaggac aaatccgccg ccctagaccg atgcataatg tgcctgtcaa
     1681 atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga
     1741 ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     1801 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
     1861 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     1921 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     1981 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     2041 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     2101 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     2161 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     2221 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     2281 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     2341 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     2401 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     2461 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     2521 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     2581 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     2641 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     2701 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     2761 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     2821 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     2881 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
     2941 tacccgtttt tttgggctag cgaattcgag ctcggtaccc ccgagcccgt agaatatgac
     3001 attaagctca taataaaagc tcaacctgac cgttagtact aacagcagaa ttactgaaac
     3061 agtagattct atcctaacga cttgtattag ttattataac ttttcaccct gtaagagaat
     3121 acactattat catgccacat tttaatcctg ttcctgtatc gaataaaaaa ttcgtctttg
     3181 atgatttcat actcaacatg gacggctccc tgctacgctc agaaaagaaa gtcaatattc
     3241 cgccaaaaga atatgccgtt ctggtcatcc tgctcgaagc cgccggcgag attgtgagta
     3301 aaaacacctt actggaccag gtatggggcg acgcggaagt taacgaagaa tctcttaccc
     3361 gctgtattta tgccttacga cgtattctgt cggaagataa agagcatcgt tacattgaaa
     3421 cactgtacgg acagggctat cggtttaatc gtccggtcgt agtggtgtct ccgccagcgc
     3481 cgcaacctac gactcataca ttggcgatac ttccttttca gatgcaggat caggttcaat
     3541 ccgagagtct gcattactct atcgtgaagg gattatcgca gtatgcgccc tttggcctga
     3601 gcgtgctgcc ggtgaccatt acgaagaact gccgcagtgt taaggatatt cttgagctca
     3661 tggatcaatt acgccccgat tattatatct ccgggcagat gatacccgat ggtaatgata
     3721 atattgtaca gattgagata gttcgggtta aaggttatca cctgctgcac caggaaagca
     3781 ttaagttgat agaacaccaa cccgcttctc tcttgcaaaa caaaattgcg aatcttttgc
     3841 tcagatgtat tcccggactt cgctgggaca caaagcagat tagcgagcta aattcgattg
     3901 acagtactat ggtttactta cgcggtaagc atgagttaaa tcaatacacc ccctatagct
     3961 tacagcaagc gcttaaattg ctgactcaat gcgttaacat gtcgccaaac agcattgcgc
     4021 cttactgtgc gctggcagaa tgctacctca gcatggcgca aatggggatt tttgataaac
     4081 aaaacgctat gatcaaagct aaagaacatg cgattaaggc gacagagctg gaccacaata
     4141 atccacaagc tttaggatta ctggggctaa ttaatacgat tcactcagaa tacatcgtcg
     4201 ggagtttgct attcaaacaa gctaacttac tttcgcccat ttctgcagat attaaatatt
     4261 attatggctg gaatcttttc atggctggtc agttggagga ggccttacaa acgattaacg
     4321 agtgtttaaa attggaccca acgcgcgcag ccgcagggat cactaagctg tggattacct
     4381 attatcatac cggtattgat gatgctatac gtttaggcga tgaattacgc tcacaacacc
     4441 tgcaggataa tccaatatta ttaagtatgc aggttatgtt tctttcgctt aaaggtaaac
     4501 atgaactggc acgaaaatta actaaagaaa tatccacgca ggaaataaca ggacttattg
     4561 ctgttaatct tctttacgct gaatattgtc agaatagtga gcgtgcctta ccgacgataa
     4621 gagaatttct ggaaagtgaa cagcgtatag ataataatcc gggattatta ccgttagtgc
     4681 tggttgccca cggcgaagct attgccgaga aaatgtggaa taaatttaaa aacgaagaca
     4741 atatttggtt caaaagatgg aaacaggatc cccgcttgat taaattacgg taaaatctga
     4801 gagaggagat atgcattatt tttttatcat cgtaatctgg ttgcttagca taaatacggc
     4861 atgggctgat tgctggcttc aggctgaaaa aatgttcaat attgaatccg aactacttta
     4921 cgctatcgcc cagcaggaat cggcgatgaa acctggcgcc attggtcata accgagatgg
     4981 ttcaaccgat cttggcctga tgcaaattaa cagcttccat atgaaaaggc tgaaaaaaat
     5041 ggggattagt gaaaaacagt tgttacagga cccctgcatt tctgtcattg tgggcgcctc
     5101 cattttatca gatatgatga aaatctacgg ttatagctgg gaggccgttg gcgcttataa
     5161 tgccgggacg tcgccgaaac gatcgaattc gagctcggta cccgggggga tcctctagag
     5221 tcgacctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     5281 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     5341 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     5401 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     5461 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     5521 agtctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6301 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     6361 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     6421 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     6481 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     6541 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     6601 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     6661 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     6721 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     6781 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     6841 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     6901 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     6961 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     7021 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     7081 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     7141 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     7201 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtctaag
     7261 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     7321 ttcacc
//
