LOCUS       sequence-296588-        5761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Pfu-RIR1-1 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5761)
  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 5761)
  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.R376
FEATURES             Location/Qualifiers
     source          1..5761
                     /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             2631..2648
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2655..2675
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2711..2742
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3253..3797
                     /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     3698..3717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3914..3931
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4413..4418,4419..4435,4436..4441)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4489..5679
                     /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(4841..4860)
                     /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 cgggggcggc tgcattgatg
     1861 gtaaagcgaa aatcatcttc gaaaacgagg gtgaagaaca cctgaccact atggaagaaa
     1921 tgtacgaacg ttataagcac ctgggagaat tttacgatga ggaatataat aggtggggaa
     1981 tcgacgtatc caatgtgccc atttatgtaa agagtttcga tccagaatca aagcgagttg
     2041 tcaaaggtaa ggttaatgta atttggaaat atgaacttgg taaagacgtt acgaaatacg
     2101 agattattac taacaaaggt acaaaaattc ttacttcgcc gtggcatcct ttttttgtcc
     2161 ttaccccgga cttcaaaatt gtagaaaaac gcgcggatga attaaaagag ggcgatattc
     2221 tcatcggatc agcgggatcc gctgctggat ctggccttca cttgatcgaa ggcttggagg
     2281 tggttaggca tatcaccacc accaacgagc cgagaacatt ttatgatttg accgttgaga
     2341 attatcagaa ttatctggcg ggtgaaaatg ggatgatctt cgttcataat acaggcttag
     2401 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2461 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2521 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2581 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag catcatcacc
     2641 atcaccacta ataatactag tagcggccgc tgcaggagtc actaagggtt agttagttag
     2701 attagcagaa agtcaaaagc ctccgaccgg aggcttttga ctaaaacttc ccttggggtt
     2761 atcattgggg ctcactcaaa ggcggtaatc agataaaaaa aatccttagc tttcgctaag
     2821 gatgatttct gctagagatg gaatagactg gatggaggcg gataaagttg caggaccact
     2881 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2941 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3001 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3061 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3121 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccttaat
     3241 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     3301 aaccgccttg cagggcggtt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     3361 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     3421 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     3481 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3541 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3601 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3661 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     3721 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     3781 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     3841 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     3901 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     3961 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     4021 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgaggt ctgcctcgtg
     4081 aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga
     4141 gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt
     4201 gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag
     4261 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     4321 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4381 caaggggtgt ttactagagg agattctcat gtttgacagc ttatcatcga taagctttaa
     4441 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat gaaatctaac
     4501 aatgcgctca tcgtcattct cggcaccgtc accctggacg ctgtaggcat aggcttggtt
     4561 atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagtat tgccagtcac
     4621 tatggcgtgc tgcttgcgct ctatgcgttg atgcaatttc tttgcgcacc cgttctcgga
     4681 gccctgtccg accgctttgg ccgccgtcca gtcctgctcg cttcgctcct tggagccact
     4741 atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc cggacgcatc
     4801 gtggcgggca tcacgggtgc cacaggtgcg gttgctggtg cctatatcgc cgacatcacc
     4861 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     4921 gtggcaggcc ccgtggccgg gggactgttg ggtgccatct ccttgcatgc accattcctt
     4981 gcggcggcgg tgctcaacgg cctcaacctc ctcctgggct gcttccttat gcaggaatcg
     5041 cataagggag agcgccgtcc gatgcccttg cgtgccttca atccagtcag ctccttccgg
     5101 tgggcgcggg gcatgactat cgtcgccgca cttatgactg ttttctttat catgcaactc
     5161 gtaggacagg ttccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     5221 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     5281 ttcgtcacgg gccccgccac caaacgtttc ggcgagaagc aggccattat cgcgggcatg
     5341 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgcggctg gatggccttc
     5401 cccattatga ttcttctcgc ttccggcggc atcggtatgc ccgcgttgca ggccatgctg
     5461 tcccgccaag tagatgacga ccatcaggga cagcttcaag ggtcgctcgc ggctcttacc
     5521 agcctcactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     5581 acatggaacg ggttggcatg gattgtaggt gccgcccttt accttgtctg cctccccgcg
     5641 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctaat aatactagct ccggcaaaaa
     5701 aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     5761 t
//
