LOCUS       sequence-333696-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB frameshift of amphipathic helix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  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 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"; journalName: "bioRxiv 
            2021.12.08.471779"
COMMENT     Sequence Label: pKP372
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      727..758
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2587..3246)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3161..3180
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3247..3349)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3741..3758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3875..4420)
                     /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(3955..3974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4640..5518)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5545..5829
                     /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     5752..5771
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atgacccttg atttacctcg ccgcttcccc tggccgacgt tactttcggt ctgcattcat
       61 ggtgctgttg tggcgggtct gctctatacc tcggtacatc aggttattga actacctgcg
      121 cctgcgcagc cgatttctgt cacgatggtt acgcctgctg atctcgaacc gccacaagcc
      181 gtccagccgc caccggagcc ggtggtagag ccagaaccgg aacctgagcc gatccccgaa
      241 ccgccaaaag aagcaccggt ggtcattgaa aagccgaagc cgaaacctaa gccaaaaccg
      301 aagccggtga aaaaggtaca ggagcagcca aaacgcgatg tcaaacccgt agagtcgcgt
      361 ccggcatcac cgtttgaaaa tacggcaccg gcacgcctga catcaagtac agcaacggct
      421 gcaaccagca agccggttac cagtgtggct tcaggaccac gcgcattaag ccgtaatcag
      481 ccgcagtatc cggcacgagc acaggcattg cgcattgaag ggcaggttaa agttaaattt
      541 gacgttacgc cggatggtcg cgtggataac gtacaaatcc tctcagccaa gcctgcgaac
      601 atgtttagcg tgaggttaaa aatgcgatgc gcagattggc gttatgagcc gggtaagcca
      661 ggcagtggga ttgtggtgaa tatcctgttt aaaattaacg gcaccaccga aattcagtaa
      721 gcagaaagtc aaaggcctcc gaccggaggc ttttgactat tactcaacag gtaaggcgcg
      781 aggttttcct tcaggatcaa ccgcgacata ctggggatcc tctagagtcg acctgcaggc
      841 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      901 taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag
      961 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     1021 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     1081 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     1141 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     1201 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     1261 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     1321 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     1381 tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg
     1441 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     1501 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag
     1561 ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg
     1621 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
     1681 cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat
     1741 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     1801 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     1861 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     1921 ccctacgtgc tgctgaagtt gcccgcaaca gagagtggaa ccaaccggtg ataccacgat
     1981 actatgactg agagtcaacg ccatgagcgg cctcatttct tattctgagt tacaacagtc
     2041 cgcaccgctg ccggtagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2101 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgcc caacagtccc
     2161 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgccctgcac cattatgttc
     2221 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     2281 aagcgctaac cgtttttatc aggctctggg aggcagaata aatgatcata tcgtcaatta
     2341 ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca
     2401 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     2461 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     2521 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     2581 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     2641 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     2701 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     2761 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     2821 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     2881 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     2941 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     3001 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     3061 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     3121 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     3181 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     3241 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3301 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     3361 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     3421 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     3481 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     3541 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     3601 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     3661 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     3721 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     3781 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     3841 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     3901 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     3961 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     4021 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     4081 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     4141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     4201 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     4261 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     4321 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     4381 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     4441 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     4501 ccccatacga tataagttgt aattctcatg tttgacagct tatcatcgat gcataatgtg
     4561 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa
     4621 ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
     4681 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     4741 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     4801 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     4861 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     4921 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     4981 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     5041 acaattgctc aagcagattt atcgccaaca gctccgaata gcgcccttcc ccttgcccgg
     5101 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     5161 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     5221 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     5281 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     5341 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     5401 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     5461 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     5521 ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc
     5581 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
     5641 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
     5701 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
     5761 catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa ctccccactg
     5821 tttctccata cccgtttttt tgggctagac ctggtttttc tactgaaatg attatgactt
     5881 ca
//
