LOCUS       sequence-337286-        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"
     terminator      742..773
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2602..3261)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3176..3195
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3262..3364)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3756..3773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3890..4435)
                     /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(3970..3989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4655..5533)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5560..5844
                     /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     5767..5786
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atgattatga cttcaatgac ccttgattta cctcgccgct tcccctggcc gacgttactt
       61 tcggtctgca ttaatggtgc tgttgtggcg ggtctgctct atacctcggt acatcaggtt
      121 attgaactac ctgcgcctgc gcagccgatt tctgtcacga tggttacgcc tgctgatctc
      181 gaaccgccac aagccgttca gccgccaccg gagccggtgg tagagccaga accggaacct
      241 gagccgatcc ccgaaccgcc aaaagaagca ccggtggtca ttgaaaagcc gaagccgaaa
      301 cctaagccaa aaccgaagcc ggtgaaaaag gtacaggagc agccaaaacg cgatgtcaaa
      361 cccgtagagt cgcgtccggc atcaccgttt gaaaatacgg caccggcacg cctgacatca
      421 agtacagcaa cggctgcaac cagcaagccg gttaccagtg tggcttcagg accacgcgca
      481 ttaagccgta atcagccgca gtatccggca cgagcacagg cattgcgcat tgaagggcag
      541 gttaaagtta aatttgacgt tacgccggat ggtcgcgtgg ataacgtaca aatcctctca
      601 gccaagcctg cgaacatgtt tgagcgtgag gtgaaaaatg cgatgcgcag atggcgttat
      661 gagccgggta agccaggcag tgggattgtg gtgaatatcc tgtttaaaat taacggcacc
      721 accgaaattc agtaagcaga aagtcaaaag cctccgaccg gaggcttttg actattactc
      781 aacaggtaag gcgcgaggtt ttccttcagg atcaaccgcg acatactggg gatcctctag
      841 agtcgacctg caggcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
      901 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaccg atgcccttga
      961 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     1021 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     1081 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     1141 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     1201 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     1261 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     1321 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     1381 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
     1441 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     1501 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     1561 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
     1621 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     1681 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     1741 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     1801 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     1861 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     1921 ggaaacgcgg aagtccccta cgtgctgctg aagttgcccg caacagagag tggaaccaac
     1981 cggtgatacc acgatactat gactgagagt caacgccatg agcggcctca tttcttattc
     2041 tgagttacaa cagtccgcac cgctgccggt agctccttcc ggtgggcgcg gggcatgact
     2101 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     2161 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgccc
     2221 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     2281 acatctgtat taacgaagcg ctaaccgttt ttatcaggct ctgggaggca gaataaatga
     2341 tcatatcgtc aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg
     2401 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     2461 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     2521 tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca
     2581 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     2641 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2701 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     2761 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     2821 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     2881 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     2941 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     3001 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
     3061 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     3121 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     3181 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     3241 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     3301 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     3361 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
     3421 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
     3481 gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag
     3541 tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg
     3601 gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact atgttggcac
     3661 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     3721 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     3781 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     3841 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     3901 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     3961 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     4021 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     4081 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     4141 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     4201 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     4261 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     4321 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     4381 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     4441 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     4501 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgtttga cagcttatca
     4561 tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     4621 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
     4681 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     4741 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     4801 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     4861 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     4921 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     4981 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     5041 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     5101 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     5161 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     5221 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     5281 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     5341 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     5401 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     5461 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     5521 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     5581 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     5641 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     5701 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     5761 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     5821 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagacctggt ttttctactg
     5881 aa
//
