LOCUS       sequence-339449-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB C18G, Q160C, F202A, W213A..
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: pKP2304
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(796..813)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      966..1052
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1144..1171
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1190..1281
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1739..2194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1826..1845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2036..2057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2750..3409)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3324..3343
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3410..3512)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3907..3924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4041..4586)
                     /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(4121..4140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4662..4681
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttctactgaa atgattatga cttcaatgac ccttgattta cctcgccgct tcccctggcc
       61 gacgttactt tcggtcggaa ttcatggtgc tgttgtggcg ggtctgctct atacctcggt
      121 acatcaggtt attgaactac ctgcgcctgc gcagccgatt tctgtcacga tggttacgcc
      181 tgctgatctc gaaccgccac aagccgttca gccgccaccg gagccggtgg tagagccaga
      241 accggaacct gagccgatcc ccgaaccgcc aaaagaagca ccggtggtca ttgaaaagcc
      301 gaagccgaaa cctaagccaa aaccgaagcc ggtgaaaaag gtacaggagc agccaaaacg
      361 cgatgtcaaa cccgtagagt cgcgtccggc atcaccgttt gaaaatacgg caccggcacg
      421 cctgacatca agtacagcaa cggctgcaac cagcaagccg gttaccagtg tggcttcagg
      481 accacgcgca ttaagccgta attgtccgca gtatccggct cgagcacagg cattgcgcat
      541 tgaagggcag gttaaagtta aatttgacgt tacgccggat ggtcgcgtgg ataacgtaca
      601 aatcctctca gccaagcctg cgaacatggc tgagcgtgag gtgaaaaatg cgatgcgcag
      661 agcgcgttat gagccgggta agccaggcag tgggattgtg gtgaatatcc tgtttaaaat
      721 taacggcacc accgaaattc agtaagcaga agcatgcaag cttggctgtt ttggcggatg
      781 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
      841 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
      901 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
      961 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1021 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1081 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     1141 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt
     1201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1261 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1321 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1381 gctgaagatc agttgggtgc agcaaactat taactggcga actacttact ctagcttccc
     1441 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1501 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1561 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1621 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1681 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     1741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1921 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     1981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2041 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
     2101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2161 gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt
     2221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     2581 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     2641 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     2701 aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc
     2761 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     2821 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     2881 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     2941 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     3001 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     3061 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     3121 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     3181 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     3241 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     3301 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     3361 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     3421 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     3481 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc
     3541 agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg
     3601 tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt
     3661 gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt
     3721 tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagtag
     3781 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     3841 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     3901 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     3961 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     4021 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     4081 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4141 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     4201 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     4261 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     4321 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     4381 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     4441 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     4501 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     4561 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     4621 aggctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     4681 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     4741 atctgctcat gtttgacagc ttatcatcga ttcagctttt cagccgccgc cagaacgtcg
     4801 tccggctgat gcctaaataa ttcgccgctg ctgttttatc gccattaaat ttctccagtg
     4861 cctgttgtgg tgtcagtaag cgtggagcgg gagttttcgc cgactcgcgc gccagttccg
     4921 gcagtagcag ctgcaaaaat tgcggcgtta aatccggcgt cggttccaca cttaaaaata
     4981 gcgccagtcg ctccatcata ttgcgcagtt cacgaatatt gcccggccag tcgtagtgca
     5041 ccagcacggt ttcgcttgcc tgtaatccct ggcgtaatgc ggcagaaaac ggggtggaga
     5101 gcgccgccag agacactttc aaaaagcttt ccgccagtgg cagaatatcc gccacccgct
     5161 cgcgcattgg tggcaattgc agacgcaaaa tactcagccg ataaaacagg tcacggcgaa
     5221 actgcccttg ccgcatatct tcttccagat tgcagtgagt ggcgctaatg acccgcacat
     5281 ccaccggaac aggctgatgc ccgccgacgc gggtgacctc tttttcttcc agcacccgca
     5341 gcagccgggt ctgcaacggc agcggcattt cgccaatctc atcgagaaac agcgtacctc
     5401 cgtgggcgat ttcaaacagc ccggcgcgac cgccgcgtcg cgagccggta aacgcccctt
     5461 cctcatagcc aaacagttct gcttccagca gcgattcggc aatcgccccg cagttgactg
     5521 caacaaacgg atgcgacttt ttgccctgtc gcgcatcgtg gcgggcaaaa tattcccgat
     5581 gaatcgcctg ggccgccagc tctttgcccg tccccgtttc cccctcaatc aacaccgctg
     5641 cactggagcg ggcatacagc aaaatagtct gccgtacttg ttccatctgt ggtgattgac
     5701 cgagcatatc gcccagcacg taacgagttc tcagggcgtt gcgggtggca tcgtgagtgt
     5761 tatggcgtaa cgacatgcgc gtcatatcca gcgcatcgct gaacgcctgg cgcacggtgg
     5821 cggcggaata gataaaaatt ccggtcattc cggcttcttc tgccagatcg gtaatcagcc
     5881 ccgcgccgac caccgcttcg gtgccgttag cttttagctc gttaatctgc ccgcgtgcgt
     5941 cttcttcggt aatgtagcta cgttgatcga ggcgcaaatt aaaggttttt tgaaacgcca
     6001 ccagtgccgg aatagtttcc tgataagtga caacgccgat agaagaggtg agttttccgg
     6061 cttttgccag tgcctgtaac acatcgtagc cgctcggttt aatcaaaata actggcactg
     6121 acaggcggct tttcaggtac gcgccgttag atccagcggc gatgatggcg tcacagcgtt
     6181 cgtttgccag tttcttgtgg atgtaggtca ccgctttttc aaagccaagc tggatagggg
     6241 taatgttcgc caggtgatca aactcgaggc tgatatcgcg aaacagctcg aacaggcgcg
     6301 ttacagatac cgtccagata accggtttgt cgtcattaag ccgtggtgga tgtgccatag
     6361 cgcaccgcaa agttaagaaa ccgaatattg ggtttagtct tgtttcataa ttgttgcaat
     6421 gaaacgcggt gaaacattgc ctgaaacgtt aactgaaacg catatttgcg gattagttca
     6481 tgactttatc tctaacaaat tgaaattaaa catttaattt tattaaggca attgtggcac
     6541 accccttgct ttgtctttat caacgcaaat aacaagttga taacaagcta gcgaattcga
     6601 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcagacctg gttt
//
