LOCUS       sequence-273515-        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the TerThyX 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  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 5746)
  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.R401
FEATURES             Location/Qualifiers
     source          1..5746
                     /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             2616..2633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2640..2660
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2696..2727
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3238..3782
                     /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     3683..3702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3899..3916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4398..4403,4404..4420,4421..4426)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4474..5664
                     /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(4826..4845)
                     /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 cattggttgt tgtttgtcgg
     1861 gcaatacaaa agttagattt aggtattcgt cttcatccca agaagcgaaa tattatgagg
     1921 aaacaattga aaaattagct aacttatggc attacgggag caaaaatcaa tatacttcta
     1981 aagatgcaaa atgtatgcaa gaaaatatat ctagccgtaa tatttttact ctggataccc
     2041 aaactaacca aattgtttct agcaaaatta ccaacatata tattaatggt gaaaaagaaa
     2101 cttacacaat aaaaactgtt tctggtaagg aaataagagc taccctagaa caccagtttt
     2161 ggactaacca agggtggaaa cgattaaaag attttaataa cagcactcaa ttatgtgaag
     2221 tgcaattagc gggatcagcc ggttcagcgg ccggttcggg aggtgttttt gtagagatag
     2281 aatctattga aaaatttggc aaagaaatta cttatgatct tgaagtcgaa cacccagaac
     2341 ataattttat agccaatggt ttagttgttc ataattcgtt cgacggcgcc ctgaagggcg
     2401 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2461 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2521 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2581 gccactccac cggcggcatg gacgagctgt acaagcatca tcaccatcac cactaataat
     2641 actagtagcg gccgctgcag gagtcactaa gggttagtta gttagattag cagaaagtca
     2701 aaagcctccg accggaggct tttgactaaa acttcccttg gggttatcat tggggctcac
     2761 tcaaaggcgg taatcagata aaaaaaatcc ttagctttcg ctaaggatga tttctgctag
     2821 agatggaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2881 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2941 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3001 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3061 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3121 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3181 cccttaacgt gagttttcgt tccactgagc gtcagacccc ttaataagat gatcttcttg
     3241 agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg
     3301 cggttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     3361 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     3421 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     3481 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     3541 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     3601 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     3661 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3721 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     3781 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     3841 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     3901 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     3961 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     4021 tagtaagcca gtatacactc cgctagcgct gaggtctgcc tcgtgaagaa ggtgttgctg
     4081 actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag ccacggttga
     4141 tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt gccacggaac
     4201 ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta
     4261 ttcaacaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     4321 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgtttact
     4381 agaggagatt ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca
     4441 cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc
     4501 attctcggca ccgtcaccct ggacgctgta ggcataggct tggttatgcc ggtactgccg
     4561 ggcctcttgc gggatatcgt ccattccgac agtattgcca gtcactatgg cgtgctgctt
     4621 gcgctctatg cgttgatgca atttctttgc gcacccgttc tcggagccct gtccgaccgc
     4681 tttggccgcc gtccagtcct gctcgcttcg ctccttggag ccactatcga ctacgcgatc
     4741 atggcgacca cacccgtcct gtggattctc tacgccggac gcatcgtggc gggcatcacg
     4801 ggtgccacag gtgcggttgc tggtgcctat atcgccgaca tcaccgatgg ggaagatcgg
     4861 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     4921 gccgggggac tgttgggtgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     4981 aacggcctca acctcctcct gggctgcttc cttatgcagg aatcgcataa gggagagcgc
     5041 cgtccgatgc ccttgcgtgc cttcaatcca gtcagctcct tccggtgggc gcggggcatg
     5101 actatcgtcg ccgcacttat gactgttttc tttatcatgc aactcgtagg acaggttccg
     5161 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     5221 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cacgggcccc
     5281 gccaccaaac gtttcggcga gaagcaggcc attatcgcgg gcatggcggc cgacgcgctg
     5341 ggctacgtct tgctggcgtt cgcgacgcgc ggctggatgg ccttccccat tatgattctt
     5401 ctcgcttccg gcggcatcgg tatgcccgcg ttgcaggcca tgctgtcccg ccaagtagat
     5461 gacgaccatc agggacagct tcaagggtcg ctcgcggctc ttaccagcct cacttcgatc
     5521 attggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg
     5581 gcatggattg taggtgccgc cctttacctt gtctgcctcc ccgcgttgcg tcgcggtgca
     5641 tggagccggg ccacctcgac ctaataatac tagctccggc aaaaaaacgg gcaaggtgtc
     5701 accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtt
//
