LOCUS       sequence-313310-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express E.coli TonB protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  AUTHORS   Larsen RA, Thomas MG, Postle K
  TITLE     Protonmotive force, ExbB and ligand-bound FepA drive conformational 
            changes in TonB.
  JOURNAL   Mol Microbiol. 1999 Mar;31(6):1809-24. doi: 
            10.1046/j.1365-2958.1999.01317.x.
  PUBMED    10209752
REFERENCE   2  (bases 1 to 5882)
  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: 
            "10.1046/j.1365-2958.1999.01317"; journalName: "Mol Microbiol"; 
            date: "1999-03"; volume: "31"; issue: "6"; pages: "1809-24"
COMMENT     Sequence Label: pKP325
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(131..148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(265..810)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(345..364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1030..1908)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1935..2219
                     /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     2142..2161
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      2999..3030
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4859..5518)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5433..5452
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5519..5621)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt
       61 catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat
      121 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
      181 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
      241 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
      301 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      361 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
      421 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
      481 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
      541 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
      601 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
      661 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
      721 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
      781 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
      841 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
      901 tataagttgt aattctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat
      961 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     1021 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     1081 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     1141 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     1201 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     1261 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     1321 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     1381 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     1441 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     1501 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     1561 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     1621 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     1681 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     1741 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     1801 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     1861 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     1921 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
     1981 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
     2041 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
     2101 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
     2161 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
     2221 cccgtttttt tgggctagac ctggtttttc tactgaaatg attatgactt caatgaccct
     2281 tgatttacct cgccgcttcc cctggccgac gttactttcg gtctgcattc atggtgctgt
     2341 tgtggcgggt ctgctctata cctcggtaca tcaggttatt gaactacctg cgcctgcgca
     2401 gccgatttct gtcacgatgg ttacgcctgc tgatctcgaa ccgccacaag ccgttcagcc
     2461 gccaccggag ccggtggtag agccagaacc ggaacctgag ccgatccccg aaccgccaaa
     2521 agaagcaccg gtggtcattg aaaagccgaa gccgaaacct aagccaaaac cgaagccggt
     2581 gaaaaaggta caggagcagc caaaacgcga tgtcaaaccc gtagagtcgc gtccggcatc
     2641 accgtttgaa aatacggcac cggcacgcct gacatcaagt acagcaacgg ctgcaaccag
     2701 caagccggtt accagtgtgg cttcaggacc acgcgcatta agccgtaatc agccgcagta
     2761 tccggcacga gcacaggcat tgcgcattga agggcaggtt aaagttaaat ttgacgttac
     2821 gccggatggt cgcgtggata acgtacaaat cctctcagcc aagcctgcga acatgtttga
     2881 gcgtgaggtg aaaaatgcga tgcgcagatg gcgttatgag ccgggtaagc caggcagtgg
     2941 gattgtggtg aatatcctgt ttaaaattaa cggcaccacc gaaattcagt aagcagaaag
     3001 tcaaaagcct ccgaccggag gcttttgact attactcaac aggtaaggcg cgaggttttc
     3061 cttcaggatc aaccgcgaca tactggggat cctctagagt cgacctgcag gcatgcacca
     3121 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     3181 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     3241 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3301 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     3361 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     3421 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     3481 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     3541 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     3601 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     3661 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     3721 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     3781 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     3841 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     3901 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     3961 gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc
     4021 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     4081 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     4141 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcccctacgt
     4201 gctgctgaag ttgcccgcaa cagagagtgg aaccaaccgg tgataccacg atactatgac
     4261 tgagagtcaa cgccatgagc ggcctcattt cttattctga gttacaacag tccgcaccgc
     4321 tgccggtagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     4381 cttctttatc atgcaactcg taggacaggt gccggcagcg cccaacagtc ccccggccac
     4441 ggggcctgcc accataccca cgccgaaaca agcgccctgc accattatgt tccggatctg
     4501 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgcta
     4561 accgttttta tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc
     4621 acggggagag cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt
     4681 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     4741 cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat
     4801 cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt
     4861 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     4921 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     4981 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     5041 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     5101 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     5161 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     5221 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     5281 cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa
     5341 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     5401 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     5461 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     5521 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5581 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     5641 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     5701 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     5761 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt
     5821 ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag
     5881 cg
//
