LOCUS       sequence-316514-        6621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TonB protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6621)
  AUTHORS   Gresock MG, Savenkova MI, Larsen RA, Ollis AA, Postle K
  TITLE     Death of the TonB Shuttle Hypothesis.
  JOURNAL   Front Microbiol. 2011 Oct 12;2:206. doi: 10.3389/fmicb.2011.00206. 
            eCollection 2011.
  PUBMED    22016747
REFERENCE   2  (bases 1 to 6621)
  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: "Front 
            Microbiol."; date: "2011-10-12"; volume: "2:206. doi"; pages: " 
            10.3389/fmicb.2011.00206. eCollection 2011"
COMMENT     Sequence Label: pKP1141
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2652..2669)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2822..2908
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3000..3027
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3046..3137
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3595..4050
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3682..3701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3892..3913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4606..5265)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5180..5199
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5266..5368)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5763..5780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5897..6442)
                     /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(5977..5996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6518..6537
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgattcag cttttcagcc gccgccagaa cgtcgtccgg ctgatgccta aataattcgc
       61 cgctgctgtt ttatcgccat taaatttctc cagtgcctgt tgtggtgtca gtaagcgtgg
      121 agcgggagtt ttcgccgact cgcgcgccag ttccggcagt agcagctgca aaaattgcgg
      181 cgttaaatcc ggcgtcggtt ccacacttaa aaatagcgcc agtcgctcca tcatattgcg
      241 cagttcacga atattgcccg gccagtcgta gtgcaccagc acggtttcgc ttgcctgtaa
      301 tccctggcgt aatgcggcag aaaacggggt ggagagcgcc gccagagaca ctttcaaaaa
      361 gctttccgcc agtggcagaa tatccgccac ccgctcgcgc agtggtggca attgcagacg
      421 caaaatactc agccgataaa acaggtcacg gcgaaactgc ccttgccgca tatcttcttc
      481 cagattgcag tgagtggcgc taatgacccg cacatccacc ggaacaggct gatgcccgcc
      541 gacgcgggtg acctcttttt cttccagcac ccgcagcagc cgggtctgca acggcagcgg
      601 catttcgcca atctcatcga gaaacagcgt acctccgtgg gcgatttcaa acagcccggc
      661 gcgaccgccg cgtcgcgagc cggtaaacgc cccttcctca tagccaaaca gttctgcttc
      721 cagcagcgat tcggcaatcg ccccgcagtt gactgcaaca aacggatgcg actttttgcc
      781 ctgtcgcgca tcgtggcggg caaaatattc ccgatgaatc gcctgggccg ccagctcttt
      841 gcccgtcccc gtttccccct caatcaacac cgctgcactg gagcgggcat acagcaaaat
      901 agtctgccgt acttgttcca tctgtggtga ttgaccgagc atatcgccca gcacgtaacg
      961 agttctcagg gcgttgcggg tggcatcgtg agtgttatgg cgtaacgaca tgcgcgtcat
     1021 atccagcgca tcgctgaacg cctggcgcac ggtggcggcg gaatagataa aaattccggt
     1081 cattccggct tcttctgcca gatcggtaat cagccccgcg ccgaccaccg cttcggtgcc
     1141 gttagctttt agctcgttaa tctgcccgcg tgcgtcttct tcggtaatgt agctacgttg
     1201 atcgaggcgc aaattaaagg ttttttgaaa cgccaccagt gccggaatag tttcctgata
     1261 agtgacaacg ccgatagaag aggtgagttt tccggctttt gccagtgcct gtaacacatc
     1321 gtagccgctc ggtttaatca aaataactgg cactgacagg cggcttttca ggtacgcgcc
     1381 gttagatcca gcggcgatga tggcgtcaca gcgttcgttt gccagtttct tgtggatgta
     1441 ggtcaccgct ttttcaaagc caagctggat aggggtaatg ttcgccaggt gatcaaactc
     1501 gaggctgata tcgcgaaaca gctcgaacag gcgcgttaca gataccgtcc agataaccgg
     1561 tttgtcgtca ttaagccgtg gtggatgtgc catagcgcac cgcaaagtta agaaaccgaa
     1621 tattgggttt agtcttgttt cataattgtt gcaatgaaac gcggtgaaac attgcctgaa
     1681 acgttaactg aaacgcatat ttgcggatta gttcatgact ttatctctaa caaattgaaa
     1741 ttaaacattt aattttatta aggcaattgt ggcacacccc ttgctttgtc tttatcaacg
     1801 caaataacaa gttgataaca agctagcgaa ttcgagctcg gtaccagacc tggtttttct
     1861 actgaaatga ttatgacttc aatgaccctt gatttacctc gccgcttccc ctggccgacg
     1921 ttactttcgg tctgcattca tggtgctgtt gtggcgggtc tgctctatac ctcggtacat
     1981 caggttattg aactacctgc gcctgcgcag ccgatttctg tcacgatggt tacgcctgct
     2041 gatctcgaac cgccacaagc cgttcagccg ccaccggagc cggtggtaga gccagaaccg
     2101 gaacctgagc cgatccccga accgccaaaa gaagcaccgg tggtcattga aaagccgaag
     2161 ccgaaaccta agccaaaacc gaagccggtg aaaaaggtac aggagcagcc aaaacgcgat
     2221 gtcaaacccg tagagtcgcg tccggcatca ccgtttgaaa atacggcacc ggcacgcctg
     2281 acatcaagta cagcaacggc tgcaaccagc aagccggtta ccagtgtggc ttcaggacca
     2341 cgcgcattaa gccgtaatca gccgcagtat ccggcacgag cacaggcatt gcgcattgaa
     2401 gggcaggtta aagttaaatt tgacgttacg ccggatggtc gcgtggataa cgtacaaatc
     2461 ctctcagcca agcctgcgaa catgtttgag cgtgaggtga aaaatgcgat gcgcagatgg
     2521 cgttatgagc cgggtaagcc aggcagtggg attgtggtga atatcctgtt taaaattaac
     2581 ggcaccaccg aaattcagta agcagaagca tgcaagcttg gctgttttgg cggatgagag
     2641 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     2701 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2761 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2821 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2881 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     2941 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3001 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     3061 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3121 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3181 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3241 aagatcagtt gggtgcagca aactattaac tggcgaacta cttactctag cttcccggca
     3301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     3601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3781 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3901 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4021 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     4081 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4141 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4201 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4261 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4321 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4381 atcctgttac cagtcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     4441 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     4501 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
     4561 gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     4621 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     4681 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     4741 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     4801 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     4861 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     4921 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     4981 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     5041 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     5101 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     5161 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     5221 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     5281 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5341 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     5401 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     5461 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
     5521 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
     5581 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagtagcgga
     5641 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     5701 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     5761 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     5821 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     5881 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     5941 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6001 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     6061 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     6121 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     6181 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     6241 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     6301 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     6361 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     6421 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctaggc
     6481 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6541 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     6601 gctcatgttt gacagcttat c
//
