LOCUS       sequence-339949-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB C18G, H20A, R212C..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  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 6654)
  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: pKP1861
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(863..880)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1033..1119
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1211..1238
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1257..1348
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1806..2261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1893..1912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2103..2124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2817..3476)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3391..3410
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3477..3579)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3974..3991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4108..4653)
                     /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(4188..4207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4729..4748
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ctagcgaatt cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcagac
       61 ctggtttttc tactgaaatg attatgactt caatgaccct tgatttacct cgccgcttcc
      121 cctggccgac gttactttcg gtcggaattg ctggtgctgt tgtggcgggt ctgctctata
      181 cctcggtaca tcaggttatt gaactacctg cgcctgcgca gccgatttct gtcacgatgg
      241 ttacgcctgc tgatctcgaa ccgccacaag ccgttcagcc gccaccggag ccggtggtag
      301 agccagaacc ggaacctgag ccgatccccg aaccgccaaa agaagcaccg gtggtcattg
      361 aaaagccgaa gccgaaacct aagccaaaac cgaagccggt gaaaaaggta caggagcagc
      421 caaaacgcga tgtcaaaccc gtagagtcgc gtccggcatc accgtttgaa aatacggcac
      481 cggcacgcct gacatcaagt acagcaacgg ctgcaaccag caagccggtt accagtgtgg
      541 cttcaggacc acgcgcatta agccgtaatc agccgcagta tccggctcga gcacaggcat
      601 tgcgcattga agggcaggtt aaagttaaat ttgacgttac gccggatggt cgcgtggata
      661 acgtacaaat cctctcagcc aagcctgcga acatgtttga gcgtgaggtg aaaaatgcga
      721 tgcgctgttg gcgttatgag ccgggtaagc caggcagtgg gattgtggtg aatatcctgt
      781 ttaaaattaa cggcaccacc gaaattcagt aagcagaagc atgcaagctt ggctgttttg
      841 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
      901 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
      961 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     1021 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1081 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1141 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1201 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     1261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1321 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1441 aaaagatgct gaagatcagt tgggtgcagc aaactattaa ctggcgaact acttactcta
     1501 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1561 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1621 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1681 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1741 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1801 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1861 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1921 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1981 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2041 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2101 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     2161 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2221 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     2281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cttacatacc
     2581 tcgctctgct aatcctgtta ccagtcaggc atttgagaag cacacggtca cactgcttcc
     2641 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     2701 tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca
     2761 cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac
     2821 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     2881 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     2941 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     3001 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     3061 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     3121 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     3181 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     3241 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     3301 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     3361 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     3421 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     3481 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     3541 tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag
     3601 ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga
     3661 tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg
     3721 atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc
     3781 ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg
     3841 ctagtagcgg agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg
     3901 cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga
     3961 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     4021 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg
     4081 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     4141 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     4201 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     4261 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     4321 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     4381 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     4441 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     4501 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     4561 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     4621 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     4681 tatttctagg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     4741 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     4801 gtagaggatc tgctcatgtt tgacagctta tcatcgattc agcttttcag ccgccgccag
     4861 aacgtcgtcc ggctgatgcc taaataattc gccgctgctg ttttatcgcc attaaatttc
     4921 tccagtgcct gttgtggtgt cagtaagcgt ggagcgggag ttttcgccga ctcgcgcgcc
     4981 agttccggca gtagcagctg caaaaattgc ggcgttaaat ccggcgtcgg ttccacactt
     5041 aaaaatagcg ccagtcgctc catcatattg cgcagttcac gaatattgcc cggccagtcg
     5101 tagtgcacca gcacggtttc gcttgcctgt aatccctggc gtaatgcggc agaaaacggg
     5161 gtggagagcg ccgccagaga cactttcaaa aagctttccg ccagtggcag aatatccgcc
     5221 acccgctcgc gcagtggtgg caattgcaga cgcaaaatac tcagccgata aaacaggtca
     5281 cggcgaaact gcccttgccg catatcttct tccagattgc agtgagtggc gctaatgacc
     5341 cgcacatcca ccggaacagg ctgatgcccg ccgacgcggg tgacctcttt ttcttccagc
     5401 acccgcagca gccgggtctg caacggcagc ggcatttcgc caatctcatc gagaaacagc
     5461 gtacctccgt gggcgatttc aaacagcccg gcgcgaccgc cgcgtcgcga gccggtaaac
     5521 gccccttcct catagccaaa cagttctgct tccagcagcg attcggcaat cgccccgcag
     5581 ttgactgcaa caaacggatg cgactttttg ccctgtcgcg catcgtggcg ggcaaaatat
     5641 tcccgatgaa tcgcctgggc cgccagctct ttgcccgtcc ccgtttcccc ctcaatcaac
     5701 accgctgcac tggagcgggc atacagcaaa atagtctgcc gtacttgttc catctgtggt
     5761 gattgaccga gcatatcgcc cagcacgtaa cgagttctca gggcgttgcg ggtggcatcg
     5821 tgagtgttat ggcgtaacga catgcgcgtc atatccagcg catcgctgaa cgcctggcgc
     5881 acggtggcgg cggaatagat aaaaattccg gtcattccgg cttcttctgc cagatcggta
     5941 atcagccccg cgccgaccac cgcttcggtg ccgttagctt ttagctcgtt aatctgcccg
     6001 cgtgcgtctt cttcggtaat gtagctacgt tgatcgaggc gcaaattaaa ggttttttga
     6061 aacgccacca gtgccggaat agtttcctga taagtgacaa cgccgataga agaggtgagt
     6121 tttccggctt ttgccagtgc ctgtaacaca tcgtagccgc tcggtttaat caaaataact
     6181 ggcactgaca ggcggctttt caggtacgcg ccgttagatc cagcggcgat gatggcgtca
     6241 cagcgttcgt ttgccagttt cttgtggatg taggtcaccg ctttttcaaa gccaagctgg
     6301 ataggggtaa tgttcgccag gtgatcaaac tcgaggctga tatcgcgaaa cagctcgaac
     6361 aggcgcgtta cagataccgt ccagataacc ggtttgtcgt cattaagccg tggtggatgt
     6421 gccatagcgc accgcaaagt taagaaaccg aatattgggt ttagtcttgt ttcataattg
     6481 ttgcaatgaa acgcggtgaa acattgcctg aaacgttaac tgaaacgcat atttgcggat
     6541 tagttcatga ctttatctct aacaaattga aattaaacat ttaattttat taaggcaatt
     6601 gtggcacacc ccttgctttg tctttatcaa cgcaaataac aagttgataa caag
//
