LOCUS       sequence-333691-        5828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB ∆(199-216).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5828)
  AUTHORS   Postle K, Kho K, Gresock M, Ghosh J, Larsen R
  TITLE     Identification of New In Vivo TonB-FepA Rendezvous Sites
  JOURNAL   bioRxiv 2021.12.08.471779
REFERENCE   2  (bases 1 to 5828)
  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: "bioRxiv 
            2021.12.08.471779"
COMMENT     Sequence Label: pKP476
FEATURES             Location/Qualifiers
     source          1..5828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      1011..1042
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2871..3530)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3445..3464
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3531..3633)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4025..4042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4159..4704)
                     /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(4239..4258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4924..5802)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagacctgg tttttctact gaaatgatta tgacttcaat gacccttgat ttacctcgcc
      361 gcttcccctg gccgacgtta ctttcggtct gcattcatgg tgctgttgtg gcgggtctgc
      421 tctatacctc ggtacatcag gttattgaac tacctgcgcc tgcgcagccg atttctgtca
      481 cgatggttac gcctgctgat ctcgaaccgc cacaagccgt tcagccgcca ccggagccgg
      541 tggtagagcc agaaccggaa cctgagccga tccccgaacc gccaaaagaa gcaccggtgg
      601 tcattgaaaa gccgaagccg aaacctaagc caaaaccgaa gccggtgaaa aaggtacagg
      661 agcagccaaa acgcgatgtc aaacccgtag agtcgcgtcc ggcatcaccg tttgaaaata
      721 cggcaccggc acgcctgaca tcaagtacag caacggctgc aaccagcaag ccggttacca
      781 gtgtggcttc aggaccacgc gcattaagcc gtaatcagcc gcagtatccg gctcgagcac
      841 aggcattgcg cattgaaggg caggttaaag ttaaatttga cgttacgccg gatggtcgcg
      901 tggataacgt acaaatcctc tcagccaagc ccccgggtaa gccaggcagt gggattgtgg
      961 tgaatatcct gtttaaaatt aacggcacca ccgaaattca gtaagcagaa agtcaaaagc
     1021 ctccgaccgg aggcttttga ctattactca acaggtaagg cgcgaggttt tccttcagga
     1081 tcaaccgcga catactgggg atcctctaga gtcgacctgc aggcatgcac cattccttgc
     1141 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     1201 taagggagag cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     1261 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     1321 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     1381 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     1441 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     1501 ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc
     1561 cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc
     1621 caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag
     1681 cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac
     1741 atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt
     1801 gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct
     1861 aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg
     1921 caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg
     1981 cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg
     2041 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     2101 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     2161 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcccctac gtgctgctga
     2221 agttgcccgc aacagagagt ggaaccaacc ggtgatacca cgatactatg actgagagtc
     2281 aacgccatga gcggcctcat ttcttattct gagttacaac agtccgcacc gctgccggta
     2341 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     2401 tcatgcaact cgtaggacag gtgccggcag cgcccaacag tcccccggcc acggggcctg
     2461 ccaccatacc cacgccgaaa caagcgccct gcaccattat gttccggatc tgcatcgcag
     2521 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc taaccgtttt
     2581 tatcaggctc tgggaggcag aataaatgat catatcgtca attattacct ccacggggag
     2641 agcctgagca aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     2701 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     2761 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     2821 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     2881 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     2941 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     3001 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     3061 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     3121 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     3181 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     3241 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     3301 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     3361 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     3421 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     3481 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     3541 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     3601 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc
     3661 cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc
     3721 gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag
     3781 tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg
     3841 ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg
     3901 gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg
     3961 gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg
     4021 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt
     4081 acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag
     4141 ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg
     4201 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4261 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     4321 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     4381 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     4441 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     4501 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     4561 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     4621 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     4681 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     4741 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
     4801 ttgtaattct catgtttgac agcttatcat cgatgcataa tgtgcctgtc aaatggacga
     4861 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
     4921 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     4981 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     5041 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     5101 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     5161 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     5221 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     5281 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     5341 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     5401 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     5461 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     5521 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     5581 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     5641 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     5701 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     5761 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     5821 attcagag
//
