LOCUS       sequence-297911-        7321 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 7321)
  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 7321)
  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..7321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1384..1449
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1462..1533
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1559..1645
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1753..2631)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2658..2942
                     /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     2865..2884
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    5181..5239
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5272..5289)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5442..5513
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6346..6440)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6461..7120)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7035..7054
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7121..7223)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7289..7307
                     /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 atcttggaaa aaaattactt gaggcggcac gcgcagggca ggatgacgag
      961 gtacgtatcc ttatggcgaa tggagccgat gtgaatgctc acgacacttt cggtttcact
     1021 cctcttcacc ttgctgcctt gtacggacac ttagagatcg tcgaggtatt attaaagaat
     1081 ggggcagacg tgaatgcgga tgactcatat ggtcgtactc ccctgcatct ggcagcgatg
     1141 cgtgggcacc ttgagattgt agaagtactg ctgaaatacg gggcagacgt aaatgccgca
     1201 gatgaggagg gacgcacacc tttgcatctt gcagcaaaac gtggccactt ggaaattgtt
     1261 gaagtactgc tgaagaacgg ggcagatgtt aacgcacagg ataaattcgg caaaacagct
     1321 ttcgatattt ccattgataa tggcaatgag gatcttgcgg aaattctgca gaagcttgtc
     1381 gacgattata aagatcatga tggcgattat aaagatcatg atattgatta taaagatgat
     1441 gatgataaat aataatctag agattataaa gatgacgatg acaaggatta taaagatgac
     1501 gatgacaagg attataaaga tgacgatgac aagtaataaa gcggccgctg attggcatca
     1561 aataaaacga aaggctcagt cgaaagactg ggactttcgt tttatctgtt gtttgtcggt
     1621 gaacgctctc ctgagtagga caaatccgcc gccctagacc gatgcataat gtgcctgtca
     1681 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     1741 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     1801 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     1861 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     1921 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     1981 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     2041 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     2101 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     2161 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     2221 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     2281 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     2341 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     2401 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     2461 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     2521 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     2581 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     2641 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     2701 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     2761 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     2821 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     2881 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     2941 atacccgttt ttttgggcta gcgaattcga gctcggtacc cgagcccgta gaatatgaca
     3001 ttaagctcat aataaaagct caacctgacc gttagtacta acagcagaat tactgaaaca
     3061 gtagattcta tcctaacgac ttgtattagt tattataact tttcaccctg taagagaata
     3121 cactattatc atgccacatt ttaatcctgt tcctgtatcg aataaaaaat tcgtctttga
     3181 tgatttcata ctcaacatgg acggctccct gctacgctca gaaaagaaag tcaatattcc
     3241 gccaaaagaa tatgccgttc tggtcatcct gctcgaagcc gccggcgaga ttgtgagtaa
     3301 aaacacctta ctggaccagg tatggggcga cgcggaagtt aacgaagaat ctcttacccg
     3361 ctgtatttat gccttacgac gtattctgtc ggaagataaa gagcatcgtt acattgaaac
     3421 actgtacgga cagggctatc ggtttaatcg tccggtcgta gtggtgtctc cgccagcgcc
     3481 gcaacctacg actcatacat tggcgatact tccttttcag atgcaggatc aggttcaatc
     3541 cgagagtctg cattactcta tcgtgaaggg attatcgcag tatgcgccct ttggcctgag
     3601 cgtgctgccg gtgaccatta cgaagaactg ccgcagtgtt aaggatattc ttgagctcat
     3661 ggatcaatta cgccccgatt attatatctc cgggcagatg atacccgatg gtaatgataa
     3721 tattgtacag attgagatag ttcgggttaa aggttatcac ctgctgcacc aggaaagcat
     3781 taagttgata gaacaccaac ccgcttctct cttgcaaaac aaaattgcga atcttttgct
     3841 cagatgtatt cccggacttc gctgggacac aaagcagatt agcgagctaa attcgattga
     3901 cagtactatg gtttacttac gcggtaagca tgagttaaat caatacaccc cctatagctt
     3961 acagcaagcg cttaaattgc tgactcaatg cgttaacatg tcgccaaaca gcattgcgcc
     4021 ttactgtgcg ctggcagaat gctacctcag catggcgcaa atggggattt ttgataaaca
     4081 aaacgctatg atcaaagcta aagaacatgc gattaaggcg acagagctgg accacaataa
     4141 tccacaagct ttaggattac tggggctaat taatacgatt cactcagaat acatcgtcgg
     4201 gagtttgcta ttcaaacaag ctaacttact ttcgcccatt tctgcagata ttaaatatta
     4261 ttatggctgg aatcttttca tggctggtca gttggaggag gccttacaaa cgattaacga
     4321 gtgtttaaaa ttggacccaa cgcgcgcagc cgcagggatc actaagctgt ggattaccta
     4381 ttatcatacc ggtattgatg atgctatacg tttaggcgat gaattacgct cacaacacct
     4441 gcaggataat ccaatattat taagtatgca ggttatgttt ctttcgctta aaggtaaaca
     4501 tgaactggca cgaaaattaa ctaaagaaat atccacgcag gaaataacag gacttattgc
     4561 tgttaatctt ctttacgctg aatattgtca gaatagtgag cgtgccttac cgacgataag
     4621 agaatttctg gaaagtgaac agcgtataga taataatccg ggattattac cgttagtgct
     4681 ggttgcccac ggcgaagcta ttgccgagaa aatgtggaat aaatttaaaa acgaagacaa
     4741 tatttggttc aaaagatgga aacaggatcc ccgcttgatt aaattacgga atctgagaga
     4801 ggagatatgc attatttttt tatcatcgta atctggttgc ttagcataaa tacggcatgg
     4861 gctgattgct ggcttcaggc tgaaaaaatg ttcaatattg aatccgaact actttacgct
     4921 atcgcccagc aggaatcggc gatgaaacct ggcgccattg gtcataaccg agatggttca
     4981 accgatcttg gcctgatgca aattaacagc ttccatatga aaaggctgaa aaaaatgggg
     5041 attagtgaaa aacagttgtt acaggacccc tgcatttctg tcattgtggg cgcctccatt
     5101 ttatcagata tgatgaaaat ctacggttat agctgggagg ccgttggcgc ttataatgcc
     5161 gggacgtcgc cgaaacgatc gaattcgagc tcggtacccg gggggatcct ctagagtcga
     5221 cctgcaggca tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     5281 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     5341 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     5401 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     5461 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagtc
     5521 tagggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa
     6361 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     6421 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     6481 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     6541 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     6601 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     6661 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     6721 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     6781 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     6841 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     6901 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     6961 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     7021 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     7081 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     7141 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     7201 tggaacctct tacgtgccga tcaacgtctc attttcgcca gatatcgacg tctaagaaac
     7261 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     7321 c
//
