LOCUS       sequence-273519-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the MP-B-DnaB 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  AUTHORS   Pinto F, Thornton EL, Wang B
  TITLE     An expanded library of orthogonal split inteins enables modular 
            multi-peptide assemblies.
  JOURNAL   Nat Commun. 2020 Mar 23;11(1):1529. doi: 10.1038/s41467-020-15272-2.
  PUBMED    32251274
REFERENCE   2  (bases 1 to 5701)
  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: "Nat 
            Commun."; date: "2020-03-23"; volume: "11(1):1529. doi"; pages: " 
            10.1038/s41467-020-15272-2"
COMMENT     Sequence Label: pFP.R368
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(48..77)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    124..145
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(146..1024)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1051..1335
                     /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     1258..1277
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1347..1358
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1510..1528)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1778..1797
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2571..2588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2595..2615
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2651..2682
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3193..3737
                     /direction=RIGHT
                     /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     3638..3657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3854..3871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4353..4358,4359..4375,4376..4381)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4429..5619
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARNFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(4781..4800)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgccacctga cgtctaagaa aaggaatatt cagcaatttg cccgtgccga agaaaggccc
       61 acccgtgaag gtgagccagt gagttgattg ctacgtaatt agttagttag cccttagtga
      121 ctcgaattcg cggccgcttc tagagttatg acaacttgac ggctacatca ttcacttttt
      181 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
      241 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
      301 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
      361 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
      421 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
      481 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
      541 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
      601 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
      661 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
      721 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
      781 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
      841 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
      901 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
      961 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     1021 ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac
     1081 attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta
     1141 ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg
     1201 tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg
     1261 ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct
     1321 ctactgtttc tccattacta gagattaaag aggagaaata ctagatggtg agcaagggcg
     1381 aagaagacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1441 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1501 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1561 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1621 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     1681 acggcggcgt ggtgaccgtg acccaggact cctccttgca ggacggcgag ttcatctaca
     1741 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     1801 tgggctggga ggcctcctcc gagcggatgt accccgagga ccaggatcag ccattggcac
     1861 taaatactga ggttcccaca ccgagtggat ggacaaccgt tggtgatctg agcgtcgggg
     1921 attatgttct cggcagtgac ggccagcccc atcgggttca gagggagact ccggtccttg
     1981 aagggcttgc tacgtatgtg gtaaggtttg acgacgggac agaaattaca gcgagtgcta
     2041 gccacggctg gaccactcaa cgtctgacgg gccatggcga ttcttacgaa accgtaactg
     2101 taaccaccga agagctggct cagactgtta caaattccaa gggacgtaaa cgtcatcgca
     2161 tccctgtagt gggtatggaa gggtcggcag gaagtgctgc tggtagtggc tcccagcatc
     2221 ggtgggttga atcagtaaca cccgttgaga gcgttccggt taaatgcatt ggtattgata
     2281 ccgaagatca tcttttccag gtgagtcgca gccgaatctt aacacacaac accaaaaatg
     2341 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2401 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2461 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2521 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag catcatcacc
     2581 atcaccacta ataatactag tagcggccgc tgcaggagtc actaagggtt agttagttag
     2641 attagcagaa agtcaaaagc ctccgaccgg aggcttttga ctaaaacttc ccttggggtt
     2701 atcattgggg ctcactcaaa ggcggtaatc agataaaaaa aatccttagc tttcgctaag
     2761 gatgatttct gctagagatg gaatagactg gatggaggcg gataaagttg caggaccact
     2821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3121 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccttaat
     3181 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     3241 aaccgccttg cagggcggtt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     3301 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     3361 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     3421 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3481 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3541 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3601 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     3661 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     3721 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     3781 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     3841 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     3901 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     3961 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgaggt ctgcctcgtg
     4021 aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga
     4081 gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt
     4141 gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag
     4201 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     4261 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4321 caaggggtgt ttactagagg agattctcat gtttgacagc ttatcatcga taagctttaa
     4381 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat gaaatctaac
     4441 aatgcgctca tcgtcattct cggcaccgtc accctggacg ctgtaggcat aggcttggtt
     4501 atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagtat tgccagtcac
     4561 tatggcgtgc tgcttgcgct ctatgcgttg atgcaatttc tttgcgcacc cgttctcgga
     4621 gccctgtccg accgctttgg ccgccgtcca gtcctgctcg cttcgctcct tggagccact
     4681 atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc cggacgcatc
     4741 gtggcgggca tcacgggtgc cacaggtgcg gttgctggtg cctatatcgc cgacatcacc
     4801 gatggggaag atcgggctcg caacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     4861 gtggcaggcc ccgtggccgg gggactgttg ggtgccatct ccttgcatgc accattcctt
     4921 gcggcggcgg tgctcaacgg cctcaacctc ctcctgggct gcttccttat gcaggaatcg
     4981 cataagggag agcgccgtcc gatgcccttg cgtgccttca atccagtcag ctccttccgg
     5041 tgggcgcggg gcatgactat cgtcgccgca cttatgactg ttttctttat catgcaactc
     5101 gtaggacagg ttccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     5161 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     5221 ttcgtcacgg gccccgccac caaacgtttc ggcgagaagc aggccattat cgcgggcatg
     5281 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgcggctg gatggccttc
     5341 cccattatga ttcttctcgc ttccggcggc atcggtatgc ccgcgttgca ggccatgctg
     5401 tcccgccaag tagatgacga ccatcaggga cagcttcaag ggtcgctcgc ggctcttacc
     5461 agcctcactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     5521 acatggaacg ggttggcatg gattgtaggt gccgcccttt accttgtctg cctccccgcg
     5581 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctaat aatactagct ccggcaaaaa
     5641 aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     5701 t
//
