LOCUS       sequence-325584-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB H20N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  AUTHORS   Swayne C, Postle K
  TITLE     Taking the Escherichia coli TonB transmembrane domain ""offline""? 
            Nonprotonatable Asn substitutes fully for TonB His20.
  JOURNAL   J Bacteriol. 2011 Aug;193(15):3693-701. doi: 10.1128/JB.05219-11. 
            Epub 2011 Jun 10.
  PUBMED    21665976
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: 
            "10.1128/JB.05219-11"; journalName: "J Bacteriol"; date: "2011-08"; 
            volume: "193"; issue: "15"; pages: "3693-701"
COMMENT     Sequence Label: pKP1193
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /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(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1613..2491)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2518..2802
                     /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     2725..2744
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      3582..3613
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(5442..5882,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc
     1561 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
     1621 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     1681 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     1741 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     1801 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     1861 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     1921 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     1981 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     2041 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     2101 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     2161 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     2221 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     2281 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     2341 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     2401 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     2461 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     2521 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     2581 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     2641 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     2701 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     2761 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta
     2821 gacctggttt ttctactgaa atgattatga cttcaatgac ccttgattta cctcgccgct
     2881 tcccctggcc gacgttactt tcggtctgca ttaatggtgc tgttgtggcg ggtctgctct
     2941 atacctcggt acatcaggtt attgaactac ctgcgcctgc gcagccgatt tctgtcacga
     3001 tggttacgcc tgctgatctc gaaccgccac aagccgttca gccgccaccg gagccggtgg
     3061 tagagccaga accggaacct gagccgatcc ccgaaccgcc aaaagaagca ccggtggtca
     3121 ttgaaaagcc gaagccgaaa cctaagccaa aaccgaagcc ggtgaaaaag gtacaggagc
     3181 agccaaaacg cgatgtcaaa cccgtagagt cgcgtccggc atcaccgttt gaaaatacgg
     3241 caccggcacg cctgacatca agtacagcaa cggctgcaac cagcaagccg gttaccagtg
     3301 tggcttcagg accacgcgca ttaagccgta atcagccgca gtatccggca cgagcacagg
     3361 cattgcgcat tgaagggcag gttaaagtta aatttgacgt tacgccggat ggtcgcgtgg
     3421 ataacgtaca aatcctctca gccaagcctg cgaacatgtt tgagcgtgag gtgaaaaatg
     3481 cgatgcgcag atggcgttat gagccgggta agccaggcag tgggattgtg gtgaatatcc
     3541 tgtttaaaat taacggcacc accgaaattc agtaagcaga aagtcaaaag cctccgaccg
     3601 gaggcttttg actattactc aacaggtaag gcgcgaggtt ttccttcagg atcaaccgcg
     3661 acatactggg gatcctctag agtcgacctg caggcatgca ccattccttg cggcggcggt
     3721 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     3781 gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3841 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3901 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3961 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     4021 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     4081 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     4141 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     4201 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     4261 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     4321 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     4381 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     4441 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     4501 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     4561 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     4621 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     4681 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     4741 ttccgtgttt cgtaaagtct ggaaacgcgg aagtccccta cgtgctgctg aagttgcccg
     4801 caacagagag tggaaccaac cggtgatacc acgatactat gactgagagt caacgccatg
     4861 agcggcctca tttcttattc tgagttacaa cagtccgcac cgctgtccgg tagctccttc
     4921 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     4981 ctcgtaggac aggtgccggc agcgcccaac agtcccccgg ccacggggcc tgccaccata
     5041 cccacgccga aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     5101 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctaaccgtt tttatcaggc
     5161 tctgggaggc agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag
     5221 caaactggcc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     5281 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg
     5341 ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta
     5401 gcaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     5461 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg
     5521 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     5581 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     5641 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     5701 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     5761 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     5821 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     5881 cg
//
