LOCUS       sequence-273509-        5671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the MP-K-gp53 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5671)
  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 5671)
  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.R369
FEATURES             Location/Qualifiers
     source          1..5671
                     /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             2541..2558
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2565..2585
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2621..2652
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3163..3707
                     /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     3608..3627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3824..3841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4323..4328,4329..4345,4346..4351)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4399..5589
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(4751..4770)
                     /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 caaccatgat tgctcatgga
     1861 caaacgcacg ggctgttacc agaagggggt ttgtacacgt tgacgatctt acaacggatg
     1921 atgaggttat gtctgttgat gatcaggggc gtacgatttg gcagcaaata gacgaagtgg
     1981 tacgcttccc atttagcgga accttatatt cactgggtgg cagagaaatt aacgccacaa
     2041 taactgccaa tcaccgcgtg gtcggactta atcgcgaaaa aactaaatgg gtggaacata
     2101 caccaacgtc actgccgggc aataagatgt gggtgtacac agctggggaa gggagtggaa
     2161 gcgccggctc ggcggccggt tcaggcttat acaagaatac tgtggaagaa gttgcatacg
     2221 aaggcgaagt ttggtgcctg agagtcccta acggtcgttt tttcattgaa gacggtggca
     2281 agattcactt aaccggaaat tcacgggcgg gcgccctgaa gggcgagatc aagcagaggc
     2341 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     2401 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     2461 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2521 gcatggacga gctgtacaag catcatcacc atcaccacta ataatactag tagcggccgc
     2581 tgcaggagtc actaagggtt agttagttag attagcagaa agtcaaaagc ctccgaccgg
     2641 aggcttttga ctaaaacttc ccttggggtt atcattgggg ctcactcaaa ggcggtaatc
     2701 agataaaaaa aatccttagc tttcgctaag gatgatttct gctagagatg gaatagactg
     2761 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2821 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2881 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2941 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3001 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3061 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3121 ttcgttccac tgagcgtcag accccttaat aagatgatct tcttgagatc gttttggtct
     3181 gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt ttcgaaggtt
     3241 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     3301 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     3361 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     3421 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     3481 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     3541 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     3601 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     3661 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     3721 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     3781 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     3841 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     3901 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     3961 cactccgcta gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct
     4021 gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt
     4081 aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg
     4141 gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccacg
     4201 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     4261 taaaactgtc tgcttacata aacagtaata caaggggtgt ttactagagg agattctcat
     4321 gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt aaattgctaa
     4381 cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcattct cggcaccgtc
     4441 accctggacg ctgtaggcat aggcttggtt atgccggtac tgccgggcct cttgcgggat
     4501 atcgtccatt ccgacagtat tgccagtcac tatggcgtgc tgcttgcgct ctatgcgttg
     4561 atgcaatttc tttgcgcacc cgttctcgga gccctgtccg accgctttgg ccgccgtcca
     4621 gtcctgctcg cttcgctcct tggagccact atcgactacg cgatcatggc gaccacaccc
     4681 gtcctgtgga ttctctacgc cggacgcatc gtggcgggca tcacgggtgc cacaggtgcg
     4741 gttgctggtg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4801 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     4861 ggtgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaacctc
     4921 ctcctgggct gcttccttat gcaggaatcg cataagggag agcgccgtcc gatgcccttg
     4981 cgtgccttca atccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     5041 cttatgactg ttttctttat catgcaactc gtaggacagg ttccggcagc gctctgggtc
     5101 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     5161 ttcggaatct tgcacgccct cgctcaagcc ttcgtcacgg gccccgccac caaacgtttc
     5221 ggcgagaagc aggccattat cgcgggcatg gcggccgacg cgctgggcta cgtcttgctg
     5281 gcgttcgcga cgcgcggctg gatggccttc cccattatga ttcttctcgc ttccggcggc
     5341 atcggtatgc ccgcgttgca ggccatgctg tcccgccaag tagatgacga ccatcaggga
     5401 cagcttcaag ggtcgctcgc ggctcttacc agcctcactt cgatcattgg accgctgatc
     5461 gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggt
     5521 gccgcccttt accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     5581 tcgacctaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
     5641 ttctttaaaa ccgaaaagat tacttcgcgt t
//
