LOCUS       sequence-339486-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TonB C18G, Q160C, F202A..
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: pKP2303
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2688..2705)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2858..2944
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3036..3063
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3082..3173
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3631..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3928..3949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4642..5301)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5216..5235
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5302..5404)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5799..5816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5933..6478)
                     /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(6013..6032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6554..6573
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcatcgatt cagcttttca gccgccgcca gaacgtcgtc cggctgatgc ctaaataatt
       61 cgccgctgct gttttatcgc cattaaattt ctccagtgcc tgttgtggtg tcagtaagcg
      121 tggagcggga gttttcgccg actcgcgcgc cagttccggc agtagcagct gcaaaaattg
      181 cggcgttaaa tccggcgtcg gttccacact taaaaatagc gccagtcgct ccatcatatt
      241 gcgcagttca cgaatattgc ccggccagtc gtagtgcacc agcacggttt cgcttgcctg
      301 taatccctgg cgtaatgcgg cagaaaacgg ggtggagagc gccgccagag acactttcaa
      361 aaagctttcc gccagtggca gaatatccgc cacccgctcg cgcagtggtg gcaattgcag
      421 acgcaaaata ctcagccgat aaaacaggtc acggcgaaac tgcccttgcc gcatatcttc
      481 ttccagattg cagtgagtgg cgctaatgac ccgcacatcc accggaacag gctgatgccc
      541 gccgacgcgg gtgacctctt tttcttccag cacccgcagc agccgggtct gcaacggcag
      601 cggcatttcg ccaatctcat cgagaaacag cgtacctccg tgggcgattt caaacagccc
      661 ggcgcgaccg ccgcgtcgcg agccggtaaa cgccccttcc tcatagccaa acagttctgc
      721 ttccagcagc gattcggcaa tcgccccgca gttgactgca acaaacggat gcgacttttt
      781 gccctgtcgc gcatcgtggc gggcaaaata ttcccgatga atcgcctggg ccgccagctc
      841 tttgcccgtc cccgtttccc cctcaatcaa caccgctgca ctggagcggg catacagcaa
      901 aatagtctgc cgtacttgtt ccatctgtgg tgattgaccg agcatatcgc ccagcacgta
      961 acgagttctc agggcgttgc gggtggcatc gtgagtgtta tggcgtaacg acatgcgcgt
     1021 catatccagc gcatcgctga acgcctggcg cacggtggcg gcggaataga taaaaattcc
     1081 ggtcattccg gcttcttctg ccagatcggt aatcagcccc gcgccgacca ccgcttcggt
     1141 gccgttagct tttagctcgt taatctgccc gcgtgcgtct tcttcggtaa tgtagctacg
     1201 ttgatcgagg cgcaaattaa aggttttttg aaacgccacc agtgccggaa tagtttcctg
     1261 ataagtgaca acgccgatag aagaggtgag ttttccggct tttgccagtg cctgtaacac
     1321 atcgtagccg ctcggtttaa tcaaaataac tggcactgac aggcggcttt tcaggtacgc
     1381 gccgttagat ccagcggcga tgatggcgtc acagcgttcg tttgccagtt tcttgtggat
     1441 gtaggtcacc gctttttcaa agccaagctg gataggggta atgttcgcca ggtgatcaaa
     1501 ctcgaggctg atatcgcgaa acagctcgaa caggcgcgtt acagataccg tccagataac
     1561 cggtttgtcg tcattaagcc gtggtggatg tgccatagcg caccgcaaag ttaagaaacc
     1621 gaatattggg tttagtcttg tttcataatt gttgcaatga aacgcggtga aacattgcct
     1681 gaaacgttaa ctgaaacgca tatttgcgga ttagttcatg actttatctc taacaaattg
     1741 aaattaaaca tttaatttta ttaaggcaat tgtggcacac cccttgcttt gtctttatca
     1801 acgcaaataa caagttgata acaagctagc gaattcgagc tcggtacccg gggatcctct
     1861 agagtcgacc tgcaggcatg cagacctggt ttttctactg aaatgattat gacttcaatg
     1921 acccttgatt tacctcgccg cttcccctgg ccgacgttac tttcggtcgg aattcatggt
     1981 gctgttgtgg cgggtctgct ctatacctcg gtacatcagg ttattgaact acctgcgcct
     2041 gcgcagccga tttctgtcac gatggttacg cctgctgatc tcgaaccgcc acaagccgtt
     2101 cagccgccac cggagccggt ggtagagcca gaaccggaac ctgagccgat ccccgaaccg
     2161 ccaaaagaag caccggtggt cattgaaaag ccgaagccga aacctaagcc aaaaccgaag
     2221 ccggtgaaaa aggtacagga gcagccaaaa cgcgatgtca aacccgtaga gtcgcgtccg
     2281 gcatcaccgt ttgaaaatac ggcaccggca cgcctgacat caagtacagc aacggctgca
     2341 accagcaagc cggttaccag tgtggcttca ggaccacgcg cattaagccg taattgtccg
     2401 cagtatccgg ctcgagcaca ggcattgcgc attgaagggc aggttaaagt taaatttgac
     2461 gttacgccgg atggtcgcgt ggataacgta caaatcctct cagccaagcc tgcgaacatg
     2521 gctgagcgtg aggtgaaaaa tgcgatgcgc agatggcgtt atgagccggg taagccaggc
     2581 agtgggattg tggtgaatat cctgtttaaa attaacggca ccaccgaaat tcagtaagca
     2641 gaagcatgca agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     2701 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     2761 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     2821 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     2881 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     2941 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     3001 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     3061 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
     3121 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3181 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3241 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcagcaaact
     3301 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3361 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3421 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3481 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3541 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3601 agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg
     3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3841 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3961 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4081 caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4321 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt caggcatttg
     4441 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     4501 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     4561 tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca
     4621 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     4681 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     4741 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     4801 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     4861 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     4921 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     4981 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     5041 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
     5101 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     5161 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     5221 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     5281 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5341 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5401 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
     5461 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
     5521 gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag
     5581 tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg
     5641 gcaaaagcac cgccggacat cagcgctagt agcggagtgt atactggctt actatgttgg
     5701 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     5761 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     5821 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     5881 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     5941 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     6001 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     6061 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     6121 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     6181 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     6241 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     6301 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     6361 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     6421 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     6481 agatcatctt attaatcaga taaaatattt ctaggctcat gagcccgaag tggcgagccc
     6541 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6601 tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca gctt
//
