LOCUS       sequence-147407-        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for arabinose-induced expression of mf-lon. Note that this 
            plasmid contains the original mf-lon cassette that is not codon 
            optimized for expression in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  AUTHORS   Cameron DE, Collins JJ
  TITLE     Tunable protein degradation in bacteria.
  JOURNAL   Nat Biotechnol. 2014 Dec;32(12):1276-81. doi: 10.1038/nbt.3053. Epub
            2014 Nov 17.
  PUBMED    25402616
REFERENCE   2  (bases 1 to 4829)
  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: "10.1038/nbt.3053"; 
            journalName: "Nat Biotechnol"; date: "2014-12"; volume: "32"; issue:
            "12"; pages: "1276-81"
COMMENT     Sequence Label: pZA16mflon
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..291
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     214..233
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             302..313
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2726..2812
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2847..2864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2981..3526)
                     /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(3061..3080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3640..3734)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3764..4555,4556..4624))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4387..4406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4625..4729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4797..4815
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
       61 ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
      121 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
      181 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
      241 tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca tagaattcat
      301 taaagaggag aaaggtacca tgagtaaaaa aataaaacta cctattttcc aaatacgcgg
      361 ttcatttatt gttcctggaa ttaaagagaa tttagaagtt ggaagaaaaa acacattagc
      421 gagcgttaat tatgcgatta aaaattcaaa taatcaaatg atcgcaatac cgcagattga
      481 tgcatctgtt gaaaaacctg aattttcaga tttacatgaa tttggaattt taattgattt
      541 tgaagtaatc aaagaatgga aagataattc attaactatt agcacaaacc ctattcaacg
      601 atgtaaagta attagctttt ttgaaaatga agatcaagtg ccgtatgcgg aagttgaatt
      661 aattgaatca attaatgatt ttagtgatga agagttaaaa gaattaattg aaaaaatctc
      721 tgatgctatc aaaactaaag ctagtttagt aacaaaacaa attaaacagt taatatcagg
      781 tgaatctgat gatttaagtt tagcttttga ttcaattatg tttaaactgg ctccgtcaaa
      841 aatattgaca aatccggaat acataacatc tccgtcatta aaaacacgat ggtcaattat
      901 tgaaaaaata atttttgctg aagatggaat cataacaaga aatgcagaat caattgatgc
      961 agcacgacaa aagaatgaaa ttgaacaaga actgaaccat aaattaaaag aaaaaatgga
     1021 taaacaacaa aaagaatatt atctgcgaga aaaaatgcga attatcaaag atgaattaga
     1081 agatgaagat gattctgatg acagttcatt agaaaaatat aaagaacgtt tagctaaaga
     1141 acctttccct gaagaagtca aacgaaaaat tatggcttct atcaaacgag ttgaagcact
     1201 tcaatcagga actcctgaat ggaatactga gaaaaattac attgattgga tgatgagtat
     1261 tccctggtgg gaagaaactg aagacttaac tgatttaaaa tatgctaaaa aaattttaga
     1321 caaacatcat tatggtatga aaaaagttaa agaaagaatt attgagtact tagctgttaa
     1381 aactaaaaca aaatcactaa aagcaccaat tattacatta gttggtcccc caggtgtagg
     1441 taaaacaagt ttagcaaaat caattgcaga agcagttgga aaaaactttg ttaaagtaag
     1501 tttaggtgga gttaaagatg aatcagaaat tcgtggacat agaaaaactt atgttggttc
     1561 aatgccagga agaattattc aaacaatgaa aagagcaaaa gtaaaaaacc cattattctt
     1621 gcttgatgaa attgataaaa tggcatctga tcacagaggt gatccagcaa gtgcaatgtt
     1681 agaggttctt gacccagaac aaaacaaaga gttttcagat cattatattg aagaaccata
     1741 tgatctaagc caagttatgt ttatagcaac tgctaactat cctgaagata ttcctgaagc
     1801 cttatatgac cgtatggaaa tcataaattt atctagctat actgaaattg aaaaagttaa
     1861 gatagcacaa gattatttag ttccaaaagc aatagaacaa catgaattaa cttcagaaga
     1921 aatatcattt actgaaggtg caattaatga aattattaaa tattacacta gagaagctgg
     1981 tgtgcgtcaa ttagaaagac atattaattc tattattaga aagtatattg ttaaaaatct
     2041 aaatggtgaa atggataaaa ttgttattga tgaaaaacaa gtaaatgatt tattaggtaa
     2101 aagaattttt gatcacactg aaaaacaaga agaatctcaa attggtgtgg ttacaggttt
     2161 agcatataca caatttggtg gagatatttt accaattgaa gtaagtttat accctggtaa
     2221 aggaaattta attcttacag gtaaacttgg tgaagttatg aaagaatctg caacaattgc
     2281 gttaacttat gttaaatcaa actttgaaaa atttggagta gataaaaaag tatttgaaga
     2341 aaatgatatt catgttcacg ttcctgaagg agctgttcca aaagatggac caagtgctgg
     2401 tatcacaatt acaactgctt taatttctgc actttcagat aaaccagttt caaaagaaat
     2461 tggaatgact ggtgaaatca cactaagagg aaatgtattg ccaattggtg gattaagaga
     2521 aaaatcaatt tctgcttcaa gaagtggatt gaaaacaatc attattccaa agaaaaatga
     2581 aagagactta gacgaaattc cagatgaagt aaaagcaaaa ctaaaaatta tacctgctga
     2641 aaaatatgaa gaggtatttg caatagtttt taaaacaaaa taataagctt gatgggggat
     2701 cccatggtac gcgtgctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2761 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     2821 ctagacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     2881 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     2941 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     3001 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     3061 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     3121 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     3181 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     3241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     3301 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     3361 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     3421 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     3481 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     3541 taatcagata aaatattact agatttcagt gcaatttatc tcttcaaatg tagcacctga
     3601 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
     3661 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     3721 tatcaacagg agtccaagcg agctcgtaaa cttggtctga cagttaccaa tgcttaatca
     3781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4741 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4801 ggcgtatcac gaggcccttt cgtcttcac
//
