LOCUS       sequence-257613-        8171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EatA expression plasmid containing 8His encoding region in 
            permissive site of eatA gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  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 8171)
  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: pTV001
FEATURES             Location/Qualifiers
     source          1..8171
                     /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             577..600
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4454..4483
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4499..4516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4572..4589)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4742..4828
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4920..4947
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4966..5057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5058..5126,5127..5918)
                     /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(5276..5295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6089..6677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6578..6597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6831..6848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6863..7003
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6929..6948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7089..7111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7267..8145)
                     /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 ataaatcatc atcatcatca tcatcatcat
      601 ctcgagatct acgataagaa tggagtgtta gttggagtgc ttgataaagc tccaatgcct
      661 gattttagta gcgccacgat gaatacaggg acattaccac caggagacca tacactgtac
      721 tcacctcaat atgttgtcac agcaaagcat gttaatggat cagatataat gagttttgga
      781 catattcaaa ataattatac tgtagtagga gagaacaacc ataatagcct tgatattaaa
      841 atacggcgtt taaataagat tgtcacggag gtcgcccctg cagaaatctc cagtgttgga
      901 gctgtaaatg gcgcttatca agaagggggg cgttttaaag ccttttatag gcttggaggt
      961 ggattgcaat atataaagga taaaaatggg aatcttacac cggtatatac aaatggtggt
     1021 ttcctaaccg gaggtactat cagtgcttta agctcatata acaatggcca aatgattacc
     1081 gcacctacag gcgatatatt taatccagct aatgggcctc ttgcaaacta tctaaataaa
     1141 ggtgatagtg gctctccttt atttgcgtat gactctctgg acaaaaaatg ggttttagtt
     1201 ggcgttctgt catcaggaag cgagcatggt aataactggg ttgtcacaac tcaagatttt
     1261 cttcatcagc aaccaaaaca tgattttgat aaaacaatat catatgactc tgaaaagggt
     1321 agcttacaat ggagatataa taaaaattca ggagtgggaa cattaagtca agagagcgtt
     1381 gtgtgggaca tgcatggaaa aaaaggggga gatctaaacg caggtaaaaa tcttcaattt
     1441 acaggaaata atggagagat tattttacac gactctatag atcaaggggc tggctatttg
     1501 cagttttttg acaactacac agttacatcc ttaactgacc aaacatggac cggaggtggt
     1561 atcattactg aaaaaggtgt aaatgtgctt tggcaggtta atggtgttaa tgatgataac
     1621 ttacataaag ttggtgaagg cacattaact gttaatggaa aaggggttaa taatggagga
     1681 ctgaaagtcg gtgatggaac cgtaattctg aatcaacgcc ctgatgataa tggacacaag
     1741 caagccttta gctctattaa catttccagt ggtcgtgcaa cagttatact ttcagatgct
     1801 aatcaagtta acccagataa aatatcatgg ggatatagag gcggtactct tgatttaaat
     1861 ggaaataatg taaactttac tcgtcttcag gctgcagatt atggtgctat tgtttctaac
     1921 aataacaaaa acaaatctga attaacactt aaattacaaa cactaaatga aaatgacatt
     1981 agtgttgatg tgaagacata tgaagttttt ggggggcatg gtagtccagg tgacttatat
     2041 tatgttcctg catcaaatac ttactttatc ctgaaatcaa aggcgtacgg tccatttttc
     2101 agtgatttag ataataccaa tgtctggcaa aatgttggtc acgatcgtga taaagcgatt
     2161 caaatcgtga aacagcagaa gattggggaa agctctcaac cttatatgtt tcatggacaa
     2221 cttaatggtt atatggatgt aaatatacat ccactctctg gtaaggatgt gctgactctt
     2281 gatggttctg ttaatctgcc cgaaggggtg ataacgaaaa agtcaggtac tctgatattt
     2341 caagggcatc cggtgattca tgctggaatg acaacctcag ccggccagag tgattgggaa
     2401 aatcgtcagt ttacaatgga taaactgagg cttgatgcag caacattcca tctctccaga
     2461 aatgctcata tgcagggaga tattagtgct gccaacggaa gcaccgtcat tctgggaagt
     2521 tctcgggtct ttactgacaa gaatgacgga accggtaatg cggcatcttc tgttgaaggg
     2581 agttccattg caacaacagc tggtgaccaa agttattaca gcggtaatgt gctgctggaa
     2641 aaccattcgt ctctagaggt cagggagaat tttactggtg gtattgaggc ttatgacagt
     2701 tctgttagtg tgacctctca gaatgctatt tttgaccatg ttggtagctt tgttaacagt
     2761 agtctgcttc tcgaaaaagg agcaaaactg acagcacaga gtggtatttt cacaaataat
     2821 actatgaaaa taaaagaaaa cgcctccctg actctgacag ggataccttc tgtaggaaag
     2881 ccagggtatt attcacctgt gacctcgact actgaaggaa ttcatctcgg tgagcgagcc
     2941 agcctttcag tgaaaaatat gggctatctg agttcaaata ttacagcaga gaactctgca
     3001 gcaattatta atctgggaga cagtaatgca actatcggga agacggactc tccattattc
     3061 agtaccttaa tgaggggata taatgctgtt ttgcagggca atattatggg gccccagagc
     3121 tcagtgaata tgaacaatgc tctgtggcac tctgatagaa attcggaact caaagagctg
     3181 aaagccaacg actcccaaat agagttgggt gtaagagggc attttgcaaa actgcgggta
     3241 aaagagctta ttgcgtctaa ctcagtgttt cttgtacatg caaacaatag ccaggctgac
     3301 cagttgaacg ttaccgacaa actgcagggc agcaacaata ctattcttgt tgactttttt
     3361 aacaaagcag ccaatggtac aaatgtgacg ttaattactg caccaaaagg cagtgatgaa
     3421 aatacattca aagccggaac ccagcagatt ggattcagta atatcacgcc agaaatcagg
     3481 acagaaaata cggatacagc cacacagtgg gtgctgactg gatatcagtc tgtcgctgat
     3541 gccagagcct cgaaaatcgc aacggacttt atggattcag gttataaatc tttcctgacg
     3601 gaagtcaata atctgaacaa acgtatggga gatttacggg atagtcaggg agatgctgga
     3661 gggtgggcgc gtatcatgaa tggtaccggt tcaggtggga gtggttacag agataactat
     3721 acccacgttc agattggtgc agacagaaag catgagctga acggtataga tttattcacc
     3781 ggtgcattac tgacttatac agacaacaat gctagcagcc aggctttcag cggtaaaaca
     3841 aaatcgctag ggggaggggt gtatgcatca ggtctctttg agtctggagc ttattttgac
     3901 ctgattggta aatatctcca tcatgataat cggtatacgt tgaattttgc ctccttgggg
     3961 gaaagaagct acacctccca ttctttgtat gctggagctg aaatcgggta tcgttatcac
     4021 atgtcagaaa atacatgggt ggaaccacag atggaactgg tttatggttc ggtatcagga
     4081 aagtcattta actggaaaga ccagggaatg caactgagta tgaaagacaa agactatcac
     4141 ccactaattg gtcgaacagg tgtggatgta ggtagagcgt tctctggaga tacctggaaa
     4201 gtaacagtac gtgcaggact gggttaccag ttcgatttgc tggcaaatgg agaaactgtt
     4261 ttacaggatg cttctggtaa aaaacacttc aaaggtgaaa aagacagcag gatgctaatg
     4321 aacgtgggga cgaatgtgga agttaaagac aatatgcgtt ttggtctgga gttggagaag
     4381 tcggcgtttg ggagatataa catagacaac tctataaatg ctaacttccg ttattatttc
     4441 tgaaagcttt ctagaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca
     4501 tcatcatcat catcattgag tttaaacggt ctccagcttg gctgttttgg cggatgagag
     4561 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     4621 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     4681 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     4741 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4801 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     4861 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     4921 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     4981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5281 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5581 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6841 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6901 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6961 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     7021 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     7081 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     7141 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga
     7201 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     7261 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     7321 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     7381 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     7441 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     7501 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     7561 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     7621 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     7681 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     7741 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     7801 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     7861 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     7921 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     7981 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     8041 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     8101 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
     8161 gccattcaga g
//
