LOCUS       sequence-339472-        8312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EatA protease expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8312)
  AUTHORS   Patel SK, Dotson J, Allen KP, Fleckenstein JM
  TITLE     Identification and molecular characterization of EatA, an 
            autotransporter protein of enterotoxigenic Escherichia coli.
  JOURNAL   Infect Immun. 2004 Mar;72(3):1786-94. doi: 
            10.1128/iai.72.3.1786-1794.2004.
  PUBMED    14977988
REFERENCE   2  (bases 1 to 8312)
  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"; doi: 
            "10.1128/iai.72.3.1786-1794"; journalName: "Infect Immun"; date: 
            "2004-03"; volume: "72"; issue: "3"; pages: "1786-94"
COMMENT     Sequence Label: pSP014
FEATURES             Location/Qualifiers
     source          1..8312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4190..4231
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4241..4258
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4314..4331)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4484..4570
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4662..4689
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4708..4799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4800..4868,4869..5660)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5018..5037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6573..6590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6605..6745
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6671..6690)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6831..6853
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7009..7887)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7914..8198
                     /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     8121..8140
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             8225..8247
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             8277..8291
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 atgaataaag tgttctctct taagtatagt tttttagcca aaggttttat tgctgtttct
       61 gaacttgccc gtcgcgtttc tgttaaaggg aaactgaaga gtgcttcatc aataattatt
      121 tcaccaataa caattgctat tgtttcttat gcacccccat ctcttgctgc aacagttaat
      181 gcagatatat cgtatcaaac atttcgggat tttgccgaaa ataaaggagc ttttatagtt
      241 ggcgcatcaa atataaatat ctacgataag aatggagtgt tagttggagt gcttgataaa
      301 gctccaatgc ctgattttag tagcgccacg atgaatacag ggacattacc accaggagac
      361 catacactgt actcacctca atatgttgtc acagcaaagc atgttaatgg atcagatata
      421 atgagttttg gacatattca aaataattat actgtagtag gagagaacaa ccataatagc
      481 cttgatatta aaatacggcg tttaaataag attgtcacgg aggtcgcccc tgcagaaatc
      541 tccagtgttg gagctgtaaa tggcgcttat caagaagggg ggcgttttaa agccttttat
      601 aggcttggag gtggattgca atatataaag gataaaaatg ggaatcttac accggtatat
      661 acaaatggtg gtttcctaac cggaggtact atcagtgctt taagctcata taacaatggc
      721 caaatgatta ccgcacctac aggcgatata tttaatccag ctaatgggcc tcttgcaaac
      781 tatctaaata aaggtgatag tggctctcct ttatttgcgt atgactctct ggacaaaaaa
      841 tgggttttag ttggcgttct gtcatcagga agcgagcatg gtaataactg ggttgtcaca
      901 actcaagatt ttcttcatca gcaaccaaaa catgattttg ataaaacaat atcatatgac
      961 tctgaaaagg gtagcttaca atggagatat aataaaaatt caggagtggg aacattaagt
     1021 caagagagcg ttgtgtggga catgcatgga aaaaaagggg gagatctaaa cgcaggtaaa
     1081 aatcttcaat ttacaggaaa taatggagag attattttac acgactctat agatcaaggg
     1141 gctggctatt tgcagttttt tgacaactac acagttacat ccttaactga ccaaacatgg
     1201 accggaggtg gtatcattac tgaaaaaggt gtaaatgtgc tttggcaggt taatggtgtt
     1261 aatgatgata acttacataa agttggtgaa ggcacattaa ctgttaatgg aaaaggggtt
     1321 aataatggag gactgaaagt cggtgatgga accgtaattc tgaatcaacg ccctgatgat
     1381 aatggacaca agcaagcctt tagctctatt aacatttcca gtggtcgtgc aacagttata
     1441 ctttcagatg ctaatcaagt taacccagat aaaatatcat ggggatatag aggcggtact
     1501 cttgatttaa atggaaataa tgtaaacttt actcgtcttc aggctgcaga ttatggtgct
     1561 attgtttcta acaataacaa aaacaaatct gaattaacac ttaaattaca aacactaaat
     1621 gaaaatgaca ttagtgttga tgtgaagaca tatgaagttt ttggggggca tggtagtcca
     1681 ggtgacttat attatgttcc tgcatcaaat acttacttta tcctgaaatc aaaggcgtac
     1741 ggtccatttt tcagtgattt agataatacc aatgtctggc aaaatgttgg tcacgatcgt
     1801 gataaagcga ttcaaatcgt gaaacagcag aagattgggg aaagctctca accttatatg
     1861 tttcatggac aacttaatgg ttatatggat gtaaatatac atccactctc tggtaaggat
     1921 gtgctgactc ttgatggttc tgttaatctg cccgaagggg tgataacgaa aaagtcaggt
     1981 actctgatat ttcaagggca tccggtgatt catgctggaa tgacaacctc agccggccag
     2041 agtgattggg aaaatcgtca gtttacaatg gataaactga ggcttgatgc agcaacattc
     2101 catctctcca gaaatgctca tatgcaggga gatattagtg ctgccaacgg aagcaccgtc
     2161 attctgggaa gttctcgggt ctttactgac aagaatgacg gaaccggtaa tgcggcatct
     2221 tctgttgaag ggagttccat tgcaacaaca gctggtgacc aaagttatta cagcggtaat
     2281 gtgctgctgg aaaaccattc gtctctagag gtcagggaga attttactgg tggtattgag
     2341 gcttatgaca gttctgttag tgtgacctct cagaatgcta tttttgacca tgttggtagc
     2401 tttgttaaca gtagtctgct tctcgaaaaa ggagcaaaac tgacagcaca gagtggtatt
     2461 ttcacaaata atactatgaa aataaaagaa aacgcctccc tgactctgac agggatacct
     2521 tctgtaggaa agccagggta ttattcacct gtgacctcga ctactgaagg aattcatctc
     2581 ggtgagcgag ccagcctttc agtgaaaaat atgggctatc tgagttcaaa tattacagca
     2641 gagaactctg cagcaattat taatctggga gacagtaatg caactatcgg gaagacggac
     2701 tctccattat tcagtacctt aatgagggga tataatgctg ttttgcaggg caatattatg
     2761 gggccccaga gctcagtgaa tatgaacaat gctctgtggc actctgatag aaattcggaa
     2821 ctcaaagagc tgaaagccaa cgactcccaa atagagttgg gtgtaagagg gcattttgca
     2881 aaactgcggg taaaagagct tattgcgtct aactcagtgt ttcttgtaca tgcaaacaat
     2941 agccaggctg accagttgaa cgttaccgac aaactgcagg gcagcaacaa tactattctt
     3001 gttgactttt ttaacaaagc agccaatggt acaaatgtga cgttaattac tgcaccaaaa
     3061 ggcagtgatg aaaatacatt caaagccgga acccagcaga ttggattcag taatatcacg
     3121 ccagaaatca ggacagaaaa tacggataca gccacacagt gggtgctgac tggatatcag
     3181 tctgtcgctg atgccagagc ctcgaaaatc gcaacggact ttatggattc aggttataaa
     3241 tctttcctga cggaagtcaa taatctgaac aaacgtatgg gagatttacg ggatagtcag
     3301 ggagatgctg gagggtgggc gcgtatcatg aatggtaccg gttcaggtgg gagtggttac
     3361 agagataact atacccacgt tcagattggt gcagacagaa agcatgagct gaacggtata
     3421 gatttattca ccggtgcatt actgacttat acagacaaca atgctagcag ccaggctttc
     3481 agcggtaaaa caaaatcgct agggggaggg gtgtatgcat caggtctctt tgagtctgga
     3541 gcttattttg acctgattgg taaatatctc catcatgata atcggtatac gttgaatttt
     3601 gcctccttgg gggaaagaag ctacacctcc cattctttgt atgctggagc tgaaatcggg
     3661 tatcgttatc acatgtcaga aaatacatgg gtggaaccac agatggaact ggtttatggt
     3721 tcggtatcag gaaagtcatt taactggaaa gaccagggaa tgcaactgag tatgaaagac
     3781 aaagactatc acccactaat tggtcgaaca ggtgtggatg taggtagagc gttctctgga
     3841 gatacctgga aagtaacagt acgtgcagga ctgggttacc agttcgattt gctggcaaat
     3901 ggagaaactg ttttacagga tgcttctggt aaaaaacact tcaaaggtga aaaagacagc
     3961 aggatgctaa tgaacgtggg gacgaatgtg gaagttaaag acaatatgcg ttttggtctg
     4021 gagttggaga agtcggcgtt tgggagatat aacatagaca actctataaa tgctaacttc
     4081 cgttattatt tctgattttg atagctgcgc agctatccaa cataatgcga gctagttaag
     4141 ggagctgata tcaccttaac gatagtacag cgacaagggc gagcttgaag gtaagcctat
     4201 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     4261 agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     4321 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     4381 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     4441 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     4501 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     4561 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     4621 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     4681 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6541 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6601 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     6661 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     6721 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     6781 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6841 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     6901 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     6961 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     7021 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     7081 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     7141 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     7201 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     7261 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     7321 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     7381 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     7441 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     7501 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     7561 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     7621 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     7681 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     7741 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     7801 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     7861 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     7921 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     7981 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     8041 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     8101 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     8161 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt gggctagaaa
     8221 taattttgtt taactttaag aaggagatat acatacccat gggctctgga tccggtgatg
     8281 acgatgacaa gctcgccctt gaggaataat aa
//
