LOCUS       sequence-273522-        5770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Ckl-Ter 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  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 5770)
  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.R363
FEATURES             Location/Qualifiers
     source          1..5770
                     /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             2640..2657
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2664..2684
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2720..2751
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3262..3806
                     /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     3707..3726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3923..3940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4422..4427,4428..4444,4445..4450)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4498..5688
                     /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(4850..4869)
                     /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 caatgggaaa gcgcttgatc
     1861 ttgacacgcc catagacacg cccacggggt ggaaaaccat gcgtgacatt gagtgtggag
     1921 actatgtttt tggagttgat ggtaaaccaa cgaaagtcat tggaacgtcg gatataatgg
     1981 ttaatcatga atgttacaaa gttacctttg aagatggaga gtttatcatc gcagataccg
     2041 agcatatttg gactgtgaca acaaaatcgt cacggaaaac cttgaaatat aagcctctta
     2101 aagggcggca acttctgcgt ccagattatc gtgaatcaaa tggatatttt gatgtaacga
     2161 ccggtgaaat ggccaagaat ttcaagcatt ttcgcaaaga cggaaagggt atcgagtata
     2221 aatacagagt accgatggcc ggcgcagttg agggtagcgc aggttcagca gcaggatcag
     2281 gtaaaaacaa atcagtaatt aacatagaaa aatacaaatc cgttcctgta aagtgtatcg
     2341 ctgtggaaga cgaaaaaaaa ttgtacctgg caggcaaaaa ccacactgcc actcacaaca
     2401 ctaccctcgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     2461 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     2521 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     2581 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagc
     2641 atcatcacca tcaccactaa taatactagt agcggccgct gcaggagtca ctaagggtta
     2701 gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc
     2761 cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct
     2821 ttcgctaagg atgatttctg ctagagatgg aatagactgg atggaggcgg ataaagttgc
     2881 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2941 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3001 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3061 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3121 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3181 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3241 ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3301 aaacgaaaaa accgccttgc agggcggttt tcgaaggttc tctgagctac caactctttg
     3361 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     3421 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     3481 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3541 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     3601 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     3661 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     3721 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     3781 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     3841 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     3901 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     3961 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     4021 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
     4081 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     4141 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     4201 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     4261 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat
     4321 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     4381 acagtaatac aaggggtgtt tactagagga gattctcatg tttgacagct tatcatcgat
     4441 aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg
     4501 aaatctaaca atgcgctcat cgtcattctc ggcaccgtca ccctggacgc tgtaggcata
     4561 ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagtatt
     4621 gccagtcact atggcgtgct gcttgcgctc tatgcgttga tgcaatttct ttgcgcaccc
     4681 gttctcggag ccctgtccga ccgctttggc cgccgtccag tcctgctcgc ttcgctcctt
     4741 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat tctctacgcc
     4801 ggacgcatcg tggcgggcat cacgggtgcc acaggtgcgg ttgctggtgc ctatatcgcc
     4861 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     4921 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gtgccatctc cttgcatgca
     4981 ccattccttg cggcggcggt gctcaacggc ctcaacctcc tcctgggctg cttccttatg
     5041 caggaatcgc ataagggaga gcgccgtccg atgcccttgc gtgccttcaa tccagtcagc
     5101 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt tttctttatc
     5161 atgcaactcg taggacaggt tccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     5221 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     5281 gctcaagcct tcgtcacggg ccccgccacc aaacgtttcg gcgagaagca ggccattatc
     5341 gcgggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgcggctgg
     5401 atggccttcc ccattatgat tcttctcgct tccggcggca tcggtatgcc cgcgttgcag
     5461 gccatgctgt cccgccaagt agatgacgac catcagggac agcttcaagg gtcgctcgcg
     5521 gctcttacca gcctcacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc
     5581 tcggcgagca catggaacgg gttggcatgg attgtaggtg ccgcccttta ccttgtctgc
     5641 ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctaata atactagctc
     5701 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     5761 acttcgcgtt
//
