LOCUS       sequence-296589-        5698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the CbP-RNR 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  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 5698)
  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.R362
FEATURES             Location/Qualifiers
     source          1..5698
                     /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             2568..2585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2592..2612
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2648..2679
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3190..3734
                     /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     3635..3654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3851..3868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4350..4355,4356..4372,4373..4378)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4426..5616
                     /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(4778..4797)
                     /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 cgctagctgt tgcttcgacg
     1861 gtcagcaaaa gacactcaca aaatcatctc atggggttaa ctacctatcc ttcaaggatc
     1921 tgtacgatac caagtataaa gataaagaac ggcgtaattt taaagttttt cacaatggaa
     1981 attgggtcga agggaagcct ataagactgc tgagaaatga taaaaaaatg tacaaaatta
     2041 cgactgtgaa taataaagaa atcttagtta cggaagatca tatcaatgta accgataaag
     2101 gagataaata caccacacaa ttaaccgaaa atgattacat agccttcaat actagaccca
     2161 caaatgccat cggctcggcg gggtcggctg ccggctcagg taccatgttt tttaagatca
     2221 aaacaataca acagtatatt tccaatgatg aatacgttta ttgttttgaa atgaaaaaca
     2281 tcgaggaacc gtactttacc ttacccaatg gtataataac gcacaattgt cgtctcggcg
     2341 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     2401 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     2461 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     2521 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagcat catcaccatc
     2581 accactaata atactagtag cggccgctgc aggagtcact aagggttagt tagttagatt
     2641 agcagaaagt caaaagcctc cgaccggagg cttttgacta aaacttccct tggggttatc
     2701 attggggctc actcaaaggc ggtaatcaga taaaaaaaat ccttagcttt cgctaaggat
     2761 gatttctgct agagatggaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2821 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2881 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2941 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3001 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3061 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3121 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccttaataag
     3181 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
     3241 cgccttgcag ggcggttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     3301 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     3361 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     3421 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     3481 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     3541 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     3601 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     3661 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     3721 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     3781 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     3841 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     3901 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     3961 tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgaggtctg cctcgtgaag
     4021 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
     4081 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
     4141 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     4201 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     4261 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     4321 ggggtgttta ctagaggaga ttctcatgtt tgacagctta tcatcgataa gctttaatgc
     4381 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat
     4441 gcgctcatcg tcattctcgg caccgtcacc ctggacgctg taggcatagg cttggttatg
     4501 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagtattgc cagtcactat
     4561 ggcgtgctgc ttgcgctcta tgcgttgatg caatttcttt gcgcacccgt tctcggagcc
     4621 ctgtccgacc gctttggccg ccgtccagtc ctgctcgctt cgctccttgg agccactatc
     4681 gactacgcga tcatggcgac cacacccgtc ctgtggattc tctacgccgg acgcatcgtg
     4741 gcgggcatca cgggtgccac aggtgcggtt gctggtgcct atatcgccga catcaccgat
     4801 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     4861 gcaggccccg tggccggggg actgttgggt gccatctcct tgcatgcacc attccttgcg
     4921 gcggcggtgc tcaacggcct caacctcctc ctgggctgct tccttatgca ggaatcgcat
     4981 aagggagagc gccgtccgat gcccttgcgt gccttcaatc cagtcagctc cttccggtgg
     5041 gcgcggggca tgactatcgt cgccgcactt atgactgttt tctttatcat gcaactcgta
     5101 ggacaggttc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     5161 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     5221 gtcacgggcc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc gggcatggcg
     5281 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gcggctggat ggccttcccc
     5341 attatgattc ttctcgcttc cggcggcatc ggtatgcccg cgttgcaggc catgctgtcc
     5401 cgccaagtag atgacgacca tcagggacag cttcaagggt cgctcgcggc tcttaccagc
     5461 ctcacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     5521 tggaacgggt tggcatggat tgtaggtgcc gccctttacc ttgtctgcct ccccgcgttg
     5581 cgtcgcggtg catggagccg ggccacctcg acctaataat actagctccg gcaaaaaaac
     5641 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgtt
//
