LOCUS       sequence-336000-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB C18G..
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: pKP1362
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(786..803)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      956..1042
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1134..1161
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1180..1271
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1729..2184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1816..1835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2026..2047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2740..3399)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3314..3333
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3400..3502)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3897..3914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4031..4576)
                     /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(4111..4130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4652..4671
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atgattatga cttcaatgac ccttgattta cctcgccgct tcccctggcc gacgttactt
       61 tcggtcggaa ttcatggtgc 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 gtatccggct 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 agcatgcaag cttggctgtt ttggcggatg agagaagatt
      781 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
      841 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
      901 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
      961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1021 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     1081 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     1141 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     1201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1381 agttgggtgc agcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1441 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1501 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1681 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     1741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1921 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     1981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2041 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     2101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2161 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     2221 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2401 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2521 ttaccagtca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     2581 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     2641 cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     2701 accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     2761 atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
     2821 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
     2881 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
     2941 gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa
     3001 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
     3061 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
     3121 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
     3181 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
     3241 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
     3301 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     3361 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     3421 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     3481 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
     3541 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
     3601 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
     3661 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
     3721 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagtag cggagtgtat
     3781 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     3841 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     3901 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     3961 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     4021 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     4081 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     4141 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     4201 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     4261 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     4321 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     4381 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     4441 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     4501 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     4561 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct aggctcatga
     4621 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     4681 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
     4741 gtttgacagc ttatcatcga ttcagctttt cagccgccgc cagaacgtcg tccggctgat
     4801 gcctaaataa ttcgccgctg ctgttttatc gccattaaat ttctccagtg cctgttgtgg
     4861 tgtcagtaag cgtggagcgg gagttttcgc cgactcgcgc gccagttccg gcagtagcag
     4921 ctgcaaaaat tgcggcgtta aatccggcgt cggttccaca cttaaaaata gcgccagtcg
     4981 ctccatcata ttgcgcagtt cacgaatatt gcccggccag tcgtagtgca ccagcacggt
     5041 ttcgcttgcc tgtaatccct ggcgtaatgc ggcagaaaac ggggtggaga gcgccgccag
     5101 agacactttc aaaaagcttt ccgccagtgg cagaatatcc gccacccgct cgcgcagtgg
     5161 tggcaattgc agacgcaaaa tactcagccg ataaaacagg tcacggcgaa actgcccttg
     5221 ccgcatatct tcttccagat tgcagtgagt ggcgctaatg acccgcacat ccaccggaac
     5281 aggctgatgc ccgccgacgc gggtgacctc tttttcttcc agcacccgca gcagccgggt
     5341 ctgcaacggc agcggcattt cgccaatctc atcgagaaac agcgtacctc cgtgggcgat
     5401 ttcaaacagc ccggcgcgac cgccgcgtcg cgagccggta aacgcccctt cctcatagcc
     5461 aaacagttct gcttccagca gcgattcggc aatcgccccg cagttgactg caacaaacgg
     5521 atgcgacttt ttgccctgtc gcgcatcgtg gcgggcaaaa tattcccgat gaatcgcctg
     5581 ggccgccagc tctttgcccg tccccgtttc cccctcaatc aacaccgctg cactggagcg
     5641 ggcatacagc aaaatagtct gccgtacttg ttccatctgt ggtgattgac cgagcatatc
     5701 gcccagcacg taacgagttc tcagggcgtt gcgggtggca tcgtgagtgt tatggcgtaa
     5761 cgacatgcgc gtcatatcca gcgcatcgct gaacgcctgg cgcacggtgg cggcggaata
     5821 gataaaaatt ccggtcattc cggcttcttc tgccagatcg gtaatcagcc ccgcgccgac
     5881 caccgcttcg gtgccgttag cttttagctc gttaatctgc ccgcgtgcgt cttcttcggt
     5941 aatgtagcta cgttgatcga ggcgcaaatt aaaggttttt tgaaacgcca ccagtgccgg
     6001 aatagtttcc tgataagtga caacgccgat agaagaggtg agttttccgg cttttgccag
     6061 tgcctgtaac acatcgtagc cgctcggttt aatcaaaata actggcactg acaggcggct
     6121 tttcaggtac gcgccgttag atccagcggc gatgatggcg tcacagcgtt cgtttgccag
     6181 tttcttgtgg atgtaggtca ccgctttttc aaagccaagc tggatagggg taatgttcgc
     6241 caggtgatca aactcgaggc tgatatcgcg aaacagctcg aacaggcgcg ttacagatac
     6301 cgtccagata accggtttgt cgtcattaag ccgtggtgga tgtgccatag cgcaccgcaa
     6361 agttaagaaa ccgaatattg ggtttagtct tgtttcataa ttgttgcaat gaaacgcggt
     6421 gaaacattgc ctgaaacgtt aactgaaacg catatttgcg gattagttca tgactttatc
     6481 tctaacaaat tgaaattaaa catttaattt tattaaggca attgtggcac accccttgct
     6541 ttgtctttat caacgcaaat aacaagttga taacaagcta gcgaattcga gctcggtacc
     6601 cggggatcct ctagagtcga cctgcaggca tgcagacctg gtttttctac tgaa
//
