LOCUS       sequence-316108-        5699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ExbB, ExbD proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5699)
  AUTHORS   Ollis AA, Manning M, Held KG, Postle K
  TITLE     Cytoplasmic membrane protonmotive force energizes periplasmic 
            interactions between ExbD and TonB.
  JOURNAL   Mol Microbiol. 2009 Aug;73(3):466-81. doi: 
            10.1111/j.1365-2958.2009.06785.x. Epub 2009 Jul 16.
  PUBMED    19627500
REFERENCE   2  (bases 1 to 5699)
  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: 
            "10.1111/j.1365-2958.2009.06785.x"; journalName: "Mol Microbiol"; 
            date: "2009-08"; volume: "73"; issue: "3"; pages: "466-81"
COMMENT     Sequence Label: pKP660
FEATURES             Location/Qualifiers
     source          1..5699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2557..2574)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2727..2813
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2905..2932
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2951..3042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3043..3111,3112..3903)
                     /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(3261..3280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3945..4400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4032..4051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4242..4263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4511..5099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5000..5019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5253..5270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5436..5458
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5596..5615
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tgggtaataa tttaatgcag acggaccttt ccgtctgggg tatgtatcag cacgccgata
     1381 ttgtcgttaa gtgcgtgatg attgggctta ttttggcctc cgtagtcacc tgggcaatct
     1441 tcttcagtaa gagcgtagag ttcttcaatc agaagcgtcg ccttaagcgc gagcagcaac
     1501 tgctggctga agcgcgttcc ttaaaccagg ccaacgatat cgccgctgat tttggtagca
     1561 aaagcttaag cctgcatttg ctcaatgaag cgcagaatga gctggaactg tcagaaggca
     1621 gcgacgataa cgaaggtatt aaagaacgta ccagcttccg tctggagcgt cgggtcgccg
     1681 cagtgggtcg tcaaatgggc cgcggtaacg gctatctggc aaccatcggc gcgatttctc
     1741 cgttcgttgg tctgtttggt acggtctggg gcatcatgaa cagctttatt ggtatcgcgc
     1801 aaacgcagac cactaacctg gcagtcgttg cgccgggtat cgcagaagct ctgttagcaa
     1861 cggcaatcgg cctcgtggca gcgattcctg cggtcgttat ctataacgta tttgcacgcc
     1921 agattggcgg ctttaaagcg atgctgggtg atgttgcagc gcaggtattg ttgctgcaaa
     1981 gccgtgacct ggatctggaa gccagcgccg ctgcgcatcc ggttcgtgtc gcacaaaaat
     2041 tacgcgcagg ataatatccg atggcaatgc atcttaacga aaacctcgac gataacggcg
     2101 aaatgcatga tatcaacgtg acgccgttta tcgacgtgat gttggttctg ctgattatct
     2161 ttatggtggc ggcaccgtta gcgacggtag atgtgaaggt gaacttgcct gcttctacca
     2221 gcacgccgca gccgcgcccg gaaaaaccgg tttatctgtc ggtgaaggca gacaactcga
     2281 tgtttatcgg taacgatccg gtcaccgatg aaacaatgat tacggcgttg aatgcgttaa
     2341 ccgaaggcaa gaaagacacc accatcttct tccgagcgga caaaaccgtc gattacgaga
     2401 cgttgatgaa ggtaatggat acgctgcatc aggcgggtta cctgaagata ggtctggtcg
     2461 gcgaagaaac cgccaaagcg aagtaagggg atcctctaga gtcgacctgc aggcatgcaa
     2521 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     2581 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     2641 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     2701 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2761 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     2821 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     2881 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     2941 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3061 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3121 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3181 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3241 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3301 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3361 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3421 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3481 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3541 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3601 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3661 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3721 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3781 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3841 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3901 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4381 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4621 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5221 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5281 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt
     5341 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5401 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5461 ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc
     5521 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5581 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     5641 ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
