LOCUS       sequence-257612-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  polyhistidine replacement of EatA passenger DUF corresponding to 
            amino acids E541 -Q617 of the native EatA protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  AUTHORS   Sheikh A, Wangdi T, Vickers TJ, Aaron B, Palmer M, Miller MJ, Kim S,
            Herring C, Simoes R, Crainic JA, Gildersleeve JC, van der Post S, 
            Hansson GC, Fleckenstein JM
  TITLE     Enterotoxigenic Escherichia coli degrades the host MUC2 mucin 
            barrier to facilitate critical pathogen-enterocyte interactions in 
            human small intestine.
  JOURNAL   Infect Immun. 2021 Nov 22:IAI0057221. doi: 10.1128/IAI.00572-21.
  PUBMED    34807735
REFERENCE   2  (bases 1 to 7946)
  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: "Infect 
            Immun. 2021 Nov 22:IAI0057221. doi: 10.1128/IAI.00572-21."
COMMENT     Sequence Label: pTW5126
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1939..1965
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             4229..4258
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4274..4291
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4347..4364)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4517..4603
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4695..4722
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4741..4832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4833..4901,4902..5693)
                     /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(5051..5070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5864..6452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6353..6372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6606..6623
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6638..6778
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6704..6723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6864..6886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7042..7920)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggataaagtg ttctctctta agtatagttt tttagccaaa
      361 ggttttattg ctgtttctga acttgcccgt cgcgtttctg ttaaagggaa actgaagagt
      421 gcttcatcaa taattatttc accaataaca attgctattg tttcttatgc acccccatct
      481 cttgctgcaa cagttaatgc agatatatcg tatcaaacat ttcgggattt tgccgaaaat
      541 aaaggagctt ttatagttgg cgcatcaaat ataaatatct acgataagaa tggagtgtta
      601 gttggagtgc ttgataaagc tccaatgcct gattttagta gcgccacgat gaatacaggg
      661 acattaccac caggagacca tacactgtac tcacctcaat atgttgtcac agcaaagcat
      721 gttaatggat cagatataat gagttttgga catattcaaa ataattatac tgtagtagga
      781 gagaacaacc ataatagcct tgatattaaa atacggcgtt taaataagat tgtcacggag
      841 gtcgcccctg cagaaatctc cagtgttgga gctgtaaatg gcgcttatca agaagggggg
      901 cgttttaaag ccttttatag gcttggaggt ggattgcaat atataaagga taaaaatggg
      961 aatcttacac cggtatatac aaatggtggt ttcctaaccg gaggtactat cagtgcttta
     1021 agctcatata acaatggcca aatgattacc gcacctacag gcgatatatt taatccagct
     1081 aatgggcctc ttgcaaacta tctaaataaa ggtgatagtg gctctccttt atttgcgtat
     1141 gactctctgg acaaaaaatg ggttttagtt ggcgttctgt catcaggaag cgagcatggt
     1201 aataactggg ttgtcacaac tcaagatttt cttcatcagc aaccaaaaca tgattttgat
     1261 aaaacaatat catatgactc tgaaaagggt agcttacaat ggagatataa taaaaattca
     1321 ggagtgggaa cattaagtca agagagcgtt gtgtgggaca tgcatggaaa aaaaggggga
     1381 gatctaaacg caggtaaaaa tcttcaattt acaggaaata atggagagat tattttacac
     1441 gactctatag atcaaggggc tggctatttg cagttttttg acaactacac agttacatcc
     1501 ttaactgacc aaacatggac cggaggtggt atcattactg aaaaaggtgt aaatgtgctt
     1561 tggcaggtta atggtgttaa tgatgataac ttacataaag ttggtgaagg cacattaact
     1621 gttaatggaa aaggggttaa taatggagga ctgaaagtcg gtgatggaac cgtaattctg
     1681 aatcaacgcc ctgatgataa tggacacaag caagccttta gctctattaa catttccagt
     1741 ggtcgtgcaa cagttatact ttcagatgct aatcaagtta acccagataa aatatcatgg
     1801 ggatatagag gcggtactct tgatttaaat ggaaataatg taaactttac tcgtcttcag
     1861 gctgcagatt atggtgctat tgtttctaac aataacaaaa acaaatctga attaacactt
     1921 aaattacaaa cactaaatca tcatcatcat catcatcatc atcatctcga gcaaccttat
     1981 atgtttcatg gacaacttaa tggttatatg gatgtaaata tacatccact ctctggtaag
     2041 gatgtgctga ctcttgatgg ttctgttaat ctgcccgaag gggtgataac gaaaaagtca
     2101 ggtactctga tatttcaagg gcatccggtg attcatgctg gaatgacaac ctcagccggc
     2161 cagagtgatt gggaaaatcg tcagtttaca atggataaac tgaggcttga tgcagcaaca
     2221 ttccatctct ccagaaatgc tcatatgcag ggagatatta gtgctgccaa cggaagcacc
     2281 gtcattctgg gaagttctcg ggtctttact gacaagaatg acggaaccgg taatgcggca
     2341 tcttctgttg aagggagttc cattgcaaca acagctggtg accaaagtta ttacagcggt
     2401 aatgtgctgc tggaaaacca ttcgtctcta gaggtcaggg agaattttac tggtggtatt
     2461 gaggcttatg acagttctgt tagtgtgacc tctcagaatg ctatttttga ccatgttggt
     2521 agctttgtta acagtagtct gcttctcgaa aaaggagcaa aactgacagc acagagtggt
     2581 attttcacaa ataatactat gaaaataaaa gaaaacgcct ccctgactct gacagggata
     2641 ccttctgtag gaaagccagg gtattattca cctgtgacct cgactactga aggaattcat
     2701 ctcggtgagc gagccagcct ttcagtgaaa aatatgggct atctgagttc aaatattaca
     2761 gcagagaact ctgcagcaat tattaatctg ggagacagta atgcaactat cgggaagacg
     2821 gactctccat tattcagtac cttaatgagg ggatataatg ctgttttgca gggcaatatt
     2881 atggggcccc agagctcagt gaatatgaac aatgctctgt ggcactctga tagaaattcg
     2941 gaactcaaag agctgaaagc caacgactcc caaatagagt tgggtgtaag agggcatttt
     3001 gcaaaactgc gggtaaaaga gcttattgcg tctaactcag tgtttcttgt acatgcaaac
     3061 aatagccagg ctgaccagtt gaacgttacc gacaaactgc agggcagcaa caatactatt
     3121 cttgttgact tttttaacaa agcagccaat ggtacaaatg tgacgttaat tactgcacca
     3181 aaaggcagtg atgaaaatac attcaaagcc ggaacccagc agattggatt cagtaatatc
     3241 acgccagaaa tcaggacaga aaatacggat acagccacac agtgggtgct gactggatat
     3301 cagtctgtcg ctgatgccag agcctcgaaa atcgcaacgg actttatgga ttcaggttat
     3361 aaatctttcc tgacggaagt caataatctg aacaaacgta tgggagattt acgggatagt
     3421 cagggagatg ctggagggtg ggcgcgtatc atgaatggta ccggttcagg tgggagtggt
     3481 tacagagata actataccca cgttcagatt ggtgcagaca gaaagcatga gctgaacggt
     3541 atagatttat tcaccggtgc attactgact tatacagaca acaatgctag cagccaggct
     3601 ttcagcggta aaacaaaatc gctaggggga ggggtgtatg catcaggtct ctttgagtct
     3661 ggagcttatt ttgacctgat tggtaaatat ctccatcatg ataatcggta tacgttgaat
     3721 tttgcctcct tgggggaaag aagctacacc tcccattctt tgtatgctgg agctgaaatc
     3781 gggtatcgtt atcacatgtc agaaaataca tgggtggaac cacagatgga actggtttat
     3841 ggttcggtat caggaaagtc atttaactgg aaagaccagg gaatgcaact gagtatgaaa
     3901 gacaaagact atcacccact aattggtcga acaggtgtgg atgtaggtag agcgttctct
     3961 ggagatacct ggaaagtaac agtacgtgca ggactgggtt accagttcga tttgctggca
     4021 aatggagaaa ctgttttaca ggatgcttct ggtaaaaaac acttcaaagg tgaaaaagac
     4081 agcaggatgc taatgaacgt ggggacgaat gtggaagtta aagacaatat gcgttttggt
     4141 ctggagttgg agaagtcggc gtttgggaga tataacatag acaactctat aaatgctaac
     4201 ttccgttatt atttctgaaa gctttctaga acaaaaactc atctcagaag aggatctgaa
     4261 tagcgccgtc gaccatcatc atcatcatca ttgagtttaa acggtctcca gcttggctgt
     4321 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     4381 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     4441 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     4501 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4561 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     4621 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     4681 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     4741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5041 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     5101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6001 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6481 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6541 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6601 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     6661 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     6721 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     6781 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6841 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6901 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     6961 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     7021 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     7081 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     7141 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     7201 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     7261 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     7321 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     7381 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     7441 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     7501 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     7561 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     7621 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     7681 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     7741 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     7801 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     7861 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     7921 acttttcata ctcccgccat tcagag
//
