LOCUS       sequence-346128-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TrpC-TonB fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  AUTHORS   Skare JT, Roof SK, Postle K
  TITLE     A mutation in the amino terminus of a hybrid TrpC-TonB protein 
            relieves overproduction lethality and results in cytoplasmic 
            accumulation.
  JOURNAL   J Bacteriol. 1989 Aug;171(8):4442-7. doi: 
            10.1128/jb.171.8.4442-4447.1989.
  PUBMED    2546922
REFERENCE   2  (bases 1 to 4863)
  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"; doi: 
            "10.1128/jb.171.8.4442-4447"; journalName: "J Bacteriol"; date: 
            "1989-08"; volume: "171"; issue: "8"; pages: "4442-7"
COMMENT     Sequence Label: pKP925
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      787..818
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(1053..1071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1139..1243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1244..1312,1313..2104)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1462..1481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2275..2863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2764..2783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3017..3034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3049..3189
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3115..3134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3275..3297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3291..3482)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgcaaaccg ttttagcgaa aatcgtcgca gacaaggcga tttgggtaga agcccgcaaa
       61 cagcagcaac cgctggaatt ccccggatcc gtcccgacgt tactttcggt ctgcattcat
      121 ggtgctgttg tggcgggtct gctctatacc tcggtacatc aggttattga actacctgcg
      181 cctgcgcagc cgatttctgt cacgatggtt acgcctgctg atctcgaacc gccacaagcc
      241 gttcagccgc caccggagcc ggtggtagag ccagaaccgg aacctgagcc gatccccgaa
      301 ccgccaaaag aagcaccggt ggtcattgaa aagccgaagc cgaaacctaa gccaaaaccg
      361 aagccggtga aaaaggtaca ggagcagcca aaacgcgatg tcaaacccgt agagtcgcgt
      421 ccggcatcac cgtttgaaaa tacggcaccg gcacgcctga catcaagtac agcaacggct
      481 gcaaccagca agccggttac cagtgtggct tcaggaccac gcgcattaag ccgtaatcag
      541 ccgcagtatc cggcacgagc acaggcattg cgcattgaag ggcaggttaa agttaaattt
      601 gacgttacgc cggatggtcg cgtggataac gtacaaatcc tctcagccaa gcctgcgaac
      661 atgtttgagc gtgaggtgaa aaatgcgatg cgcagatggc gttatgagcc gggtaagcca
      721 ggcagtggga ttgtggtgaa tatcctgttt aaaattaacg gcaccaccga aattcagtaa
      781 gcagaaagtc aaaagcctcc gaccggaggc ttttgactat tactcaacag gtaaggcgcg
      841 aggttttcct tcaggatcaa ccgcgacata cttaaataat gcttctgtcg ctttatagcg
      901 ttgcccaatt ggttcagacg ctactttttt cacccacact tcaatattaa tgctgaccga
      961 tgtcgtccct ttctggacac agcgtgcata gcagcacacc acatcgccga ccgcaaccgg
     1021 gacggatccg gggaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     1081 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1141 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1201 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1261 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1321 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1381 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1441 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     1501 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1561 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1621 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1681 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1741 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     1801 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1861 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1921 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1981 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2041 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2101 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2401 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2881 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2941 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3001 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3061 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     3121 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3181 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3241 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3301 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     3361 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     3421 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     3481 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     3541 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     3601 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     3661 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     3721 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3781 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3841 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3901 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca
     3961 acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc
     4021 tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt
     4081 ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc
     4141 tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc
     4201 atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc
     4261 caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt
     4321 cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag
     4381 cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt
     4441 agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     4501 tggcgggacc agtgacgaag gcttgagcat tcaacgccag tcccgaacgt gaaatttcct
     4561 ctcttgctgg cgcgattgca gctgtggtgt catggtcggt gatcgccagg gtgccgacgc
     4621 gcatctcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     4681 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     4741 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     4801 gttgacaatt aatcatcgaa ctagttaact agtacgcaag ttcacgtaaa aagggtatcg
     4861 acg
//
