LOCUS       sequence-306304-        7323 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 7323)
  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 7323)
  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..7323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1380..1445
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1458..1529
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      1555..1641
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1749..2627)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2654..2938
                     /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     2861..2880
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    5182..5240
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5273..5290)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5443..5514
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5671..6259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5751..5770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6347..6441)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6462..7121)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7036..7055
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7122..7224)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     7290..7308
                     /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 cctgggtaag aaactgctgg aagctgctcg tgctggtcag gacgacgaag
      961 ttcgtatcct gatggctaac ggtgctgacg ttaacgctac tgacaatgat ggttatactc
     1021 cgctgcacct ggctgcttct aatggtcacc tggaaatcgt tgaagttctg ctgaagaacg
     1081 gtgctgacgt taacgcttct gaccttactg gtattactcc gctgcacctg gctgctgcta
     1141 ctggtcacct ggaaatcgtt gaagttctgc tgaagcacgg tgctgacgtt aacgcttatg
     1201 acaatgatgg tcatactccg ctgcacctgg ctgctaagta tggtcacctg gaaatcgttg
     1261 aagttctgct gaagcacggt gctgacgtta acgctcagga caaattcggt aagaccgctt
     1321 tcgacatctc catcgacaac ggtaacgagg acctggctga aatcctgcaa aaggtcgacg
     1381 attataaaga tcatgatggc gattataaag atcatgatat tgattataaa gatgatgatg
     1441 ataaataata atctagagat tataaagatg acgatgacaa ggattataaa gatgacgatg
     1501 acaaggatta taaagatgac gatgacaagt aataaagcgg ccgctgattg gcatcaaata
     1561 aaacgaaagg ctcagtcgaa agactgggac tttcgtttta tctgttgttt gtcggtgaac
     1621 gctctcctga gtaggacaaa tccgccgccc tagaccgatg cataatgtgc ctgtcaaatg
     1681 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     1741 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     1801 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     1861 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     1921 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     1981 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     2041 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     2101 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     2161 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     2221 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     2281 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     2341 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     2401 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     2461 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     2521 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     2581 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     2641 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     2701 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     2761 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     2821 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     2881 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     2941 ccgttttttt gggctagcga attcgagctc ggtacccccg agcccgtaga atatgacatt
     3001 aagctcataa taaaagctca acctgaccgt tagtactaac agcagaatta ctgaaacagt
     3061 agattctatc ctaacgactt gtattagtta ttataacttt tcaccctgta agagaataca
     3121 ctattatcat gccacatttt aatcctgttc ctgtatcgaa taaaaaattc gtctttgatg
     3181 atttcatact caacatggac ggctccctgc tacgctcaga aaagaaagtc aatattccgc
     3241 caaaagaata tgccgttctg gtcatcctgc tcgaagccgc cggcgagatt gtgagtaaaa
     3301 acaccttact ggaccaggta tggggcgacg cggaagttaa cgaagaatct cttacccgct
     3361 gtatttatgc cttacgacgt attctgtcgg aagataaaga gcatcgttac attgaaacac
     3421 tgtacggaca gggctatcgg tttaatcgtc cggtcgtagt ggtgtctccg ccagcgccgc
     3481 aacctacgac tcatacattg gcgatacttc cttttcagat gcaggatcag gttcaatccg
     3541 agagtctgca ttactctatc gtgaagggat tatcgcagta tgcgcccttt ggcctgagcg
     3601 tgctgccggt gaccattacg aagaactgcc gcagtgttaa ggatattctt gagctcatgg
     3661 atcaattacg ccccgattat tatatctccg ggcagatgat acccgatggt aatgataata
     3721 ttgtacagat tgagatagtt cgggttaaag gttatcacct gctgcaccag gaaagcatta
     3781 agttgataga acaccaaccc gcttctctct tgcaaaacaa aattgcgaat cttttgctca
     3841 gatgtattcc cggacttcgc tgggacacaa agcagattag cgagctaaat tcgattgaca
     3901 gtactatggt ttacttacgc ggtaagcatg agttaaatca atacaccccc tatagcttac
     3961 agcaagcgct taaattgctg actcaatgcg ttaacatgtc gccaaacagc attgcgcctt
     4021 actgtgcgct ggcagaatgc tacctcagca tggcgcaaat ggggattttt gataaacaaa
     4081 acgctatgat caaagctaaa gaacatgcga ttaaggcgac agagctggac cacaataatc
     4141 cacaagcttt aggattactg gggctaatta atacgattca ctcagaatac atcgtcggga
     4201 gtttgctatt caaacaagct aacttacttt cgcccatttc tgcagatatt aaatattatt
     4261 atggctggaa tcttttcatg gctggtcagt tggaggaggc cttacaaacg attaacgagt
     4321 gtttaaaatt ggacccaacg cgcgcagccg cagggatcac taagctgtgg attacctatt
     4381 atcataccgg tattgatgat gctatacgtt taggcgatga attacgctca caacacctgc
     4441 aggataatcc aatattatta agtatgcagg ttatgtttct ttcgcttaaa ggtaaacatg
     4501 aactggcacg aaaattaact aaagaaatat ccacgcagga aataacagga cttattgctg
     4561 ttaatcttct ttacgctgaa tattgtcaga atagtgagcg tgccttaccg acgataagag
     4621 aatttctgga aagtgaacag cgtatagata ataatccggg attattaccg ttagtgctgg
     4681 ttgcccacgg cgaagctatt gccgagaaaa tgtggaataa atttaaaaac gaagacaata
     4741 tttggttcaa aagatggaaa caggatcccc gcttgattaa attacggtaa aatctgagag
     4801 aggagatatg cattattttt ttatcatcgt aatctggttg cttagcataa atacggcatg
     4861 ggctgattgc tggcttcagg ctgaaaaaat gttcaatatt gaatccgaac tactttacgc
     4921 tatcgcccag caggaatcgg cgatgaaacc tggcgccatt ggtcataacc gagatggttc
     4981 aaccgatctt ggcctgatgc aaattaacag cttccatatg aaaaggctga aaaaaatggg
     5041 gattagtgaa aaacagttgt tacaggaccc ctgcatttct gtcattgtgg gcgcctccat
     5101 tttatcagat atgatgaaaa tctacggtta tagctgggag gccgttggcg cttataatgc
     5161 cgggacgtcg ccgaaacgat cgaattcgag ctcggtaccc ggggggatcc tctagagtcg
     5221 acctgcaggc atgcaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     5281 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     5341 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     5401 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     5461 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5521 ctagggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5581 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5641 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5701 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5761 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5821 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5881 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5941 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6001 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6061 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6121 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6181 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6241 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6301 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
     6361 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     6421 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     6481 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     6541 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     6601 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     6661 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     6721 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     6781 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     6841 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     6901 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     6961 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     7021 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     7081 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     7141 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     7201 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtctaagaaa
     7261 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     7321 acc
//
