LOCUS       sequence-273520-        5710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Cro-RIR1 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5710)
  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 5710)
  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.R355
FEATURES             Location/Qualifiers
     source          1..5710
                     /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             2580..2597
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2604..2624
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2660..2691
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3202..3746
                     /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     3647..3666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3863..3880
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4362..4367,4368..4384,4385..4390)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4438..5628
                     /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(4790..4809)
                     /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 cgtgaatcag tgctttacgc
     1861 ccgacacacc aatttttacg aacgatggct ttgtatcgat tgaaaacata aagcctcata
     1921 tgaaggttat gacctctgac ggtacgttcc gcaatgttaa taaaattttt aaaaacaacg
     1981 tcaataagaa tattctaaag atcaatacga ctcactcatt agaagaaatc aaatgtacaa
     2041 aagagcatga tattttaatc tatcagaata ttaataatga atctaactac gaacaaatta
     2101 cacattatat tgaaacgaac aaatatactc ctaactttgt aaaagcatcc gagttaaagg
     2161 taggtgattt catggtaatc ccgaaaatcc agattgggtc cgcggggagc gcggccggta
     2221 gcggatgcat actgacgaag atcgaaaaca ttgatatgtg tgagtactcc ggctatgtct
     2281 atgatttgaa catcgaagaa aaccacaatt atttaacctc ctcgggtatt gttcataact
     2341 ccgggagggg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     2401 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     2461 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     2521 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagc
     2581 atcatcacca tcaccactaa taatactagt agcggccgct gcaggagtca ctaagggtta
     2641 gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc
     2701 cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct
     2761 ttcgctaagg atgatttctg ctagagatgg aatagactgg atggaggcgg ataaagttgc
     2821 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2881 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2941 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3001 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3061 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3121 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3181 ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3241 aaacgaaaaa accgccttgc agggcggttt tcgaaggttc tctgagctac caactctttg
     3301 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     3361 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     3421 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3481 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     3541 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     3601 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     3661 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     3721 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     3781 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     3841 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     3901 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     3961 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
     4021 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     4081 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     4141 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     4201 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat
     4261 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     4321 acagtaatac aaggggtgtt tactagagga gattctcatg tttgacagct tatcatcgat
     4381 aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg
     4441 aaatctaaca atgcgctcat cgtcattctc ggcaccgtca ccctggacgc tgtaggcata
     4501 ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagtatt
     4561 gccagtcact atggcgtgct gcttgcgctc tatgcgttga tgcaatttct ttgcgcaccc
     4621 gttctcggag ccctgtccga ccgctttggc cgccgtccag tcctgctcgc ttcgctcctt
     4681 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat tctctacgcc
     4741 ggacgcatcg tggcgggcat cacgggtgcc acaggtgcgg ttgctggtgc ctatatcgcc
     4801 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     4861 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gtgccatctc cttgcatgca
     4921 ccattccttg cggcggcggt gctcaacggc ctcaacctcc tcctgggctg cttccttatg
     4981 caggaatcgc ataagggaga gcgccgtccg atgcccttgc gtgccttcaa tccagtcagc
     5041 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt tttctttatc
     5101 atgcaactcg taggacaggt tccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     5161 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     5221 gctcaagcct tcgtcacggg ccccgccacc aaacgtttcg gcgagaagca ggccattatc
     5281 gcgggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgcggctgg
     5341 atggccttcc ccattatgat tcttctcgct tccggcggca tcggtatgcc cgcgttgcag
     5401 gccatgctgt cccgccaagt agatgacgac catcagggac agcttcaagg gtcgctcgcg
     5461 gctcttacca gcctcacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc
     5521 tcggcgagca catggaacgg gttggcatgg attgtaggtg ccgcccttta ccttgtctgc
     5581 ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctaata atactagctc
     5641 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     5701 acttcgcgtt
//
