LOCUS       sequence-273508-        5704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the CIV-RIR1 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5704)
  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 5704)
  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.R358
FEATURES             Location/Qualifiers
     source          1..5704
                     /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             2574..2591
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2598..2618
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2654..2685
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3196..3740
                     /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     3641..3660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3857..3874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4356..4361,4362..4378,4379..4384)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4432..5622
                     /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(4784..4803)
                     /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 ctcgaatctc tgcgttgcac
     1861 ccgaaaccat gatcttgacc gaagacggtc agtttccgat taaggacctg gaggggaaga
     1921 taattaaggt ttggaatggc aacgaattta gctctgtcac agttgtaaag actggcactg
     1981 agaaagagct gctggaggtg gagttgagta atggttgtac attaagctgt actccggaac
     2041 ataaatttat cattgtaaaa agttacaccg aagcaaaaaa acaaaaaacc gatgataacg
     2101 cgatcgccaa tgccgaacgc gtagacgcgc aggatttgaa acctagaatg aagctaatca
     2161 aattcgatct gccgacgctg tttggtaata gcgggagtgc gggcagcgca gcaggttcgg
     2221 gacgttttgt tgaggtcctg aaggtgaata aaacgggtcg agtcgacgat acgtattgct
     2281 ttacggaacc gattaatcat gcgggggtgt ttaacgggat cctgaccggt caatgctcag
     2341 agggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2401 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2461 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2521 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagcatcatc
     2581 accatcacca ctaataatac tagtagcggc cgctgcagga gtcactaagg gttagttagt
     2641 tagattagca gaaagtcaaa agcctccgac cggaggcttt tgactaaaac ttcccttggg
     2701 gttatcattg gggctcactc aaaggcggta atcagataaa aaaaatcctt agctttcgct
     2761 aaggatgatt tctgctagag atggaataga ctggatggag gcggataaag ttgcaggacc
     2821 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2881 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2941 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3001 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3061 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3121 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagacccctt
     3181 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     3241 aaaaaccgcc ttgcagggcg gttttcgaag gttctctgag ctaccaactc tttgaaccga
     3301 ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg
     3361 gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc
     3421 ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3481 ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact
     3541 gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc
     3601 gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt
     3661 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     3721 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat
     3781 cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag
     3841 tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat
     3901 attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca
     3961 ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgctga ggtctgcctc
     4021 gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc agccagaaag
     4081 tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg attttgaact
     4141 tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact
     4201 cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca
     4261 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     4321 atacaagggg tgtttactag aggagattct catgtttgac agcttatcat cgataagctt
     4381 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
     4441 aacaatgcgc tcatcgtcat tctcggcacc gtcaccctgg acgctgtagg cataggcttg
     4501 gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag tattgccagt
     4561 cactatggcg tgctgcttgc gctctatgcg ttgatgcaat ttctttgcgc acccgttctc
     4621 ggagccctgt ccgaccgctt tggccgccgt ccagtcctgc tcgcttcgct ccttggagcc
     4681 actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggattctcta cgccggacgc
     4741 atcgtggcgg gcatcacggg tgccacaggt gcggttgctg gtgcctatat cgccgacatc
     4801 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     4861 atggtggcag gccccgtggc cgggggactg ttgggtgcca tctccttgca tgcaccattc
     4921 cttgcggcgg cggtgctcaa cggcctcaac ctcctcctgg gctgcttcct tatgcaggaa
     4981 tcgcataagg gagagcgccg tccgatgccc ttgcgtgcct tcaatccagt cagctccttc
     5041 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgttttctt tatcatgcaa
     5101 ctcgtaggac aggttccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     5161 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     5221 gccttcgtca cgggccccgc caccaaacgt ttcggcgaga agcaggccat tatcgcgggc
     5281 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgcgg ctggatggcc
     5341 ttccccatta tgattcttct cgcttccggc ggcatcggta tgcccgcgtt gcaggccatg
     5401 ctgtcccgcc aagtagatga cgaccatcag ggacagcttc aagggtcgct cgcggctctt
     5461 accagcctca cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     5521 agcacatgga acgggttggc atggattgta ggtgccgccc tttaccttgt ctgcctcccc
     5581 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct aataatacta gctccggcaa
     5641 aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
     5701 cgtt
//
