LOCUS       sequence-296599-        5662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the chimeric 
            Npu/SspDnaE intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5662)
  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 5662)
  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.R400
FEATURES             Location/Qualifiers
     source          1..5662
                     /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             2532..2549
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2556..2576
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2612..2643
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3154..3698
                     /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     3599..3618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3815..3832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4314..4319,4320..4336,4337..4342)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4390..5580
                     /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(4742..4761)
                     /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 cggcggaaaa tgcctgagct
     1861 atgaaaccga aattctgacc gtggaatatg gcagcctgcc gattggcaaa attgtggaaa
     1921 aacgcattga atgcaccgtg tatagcgtgg ataacaacgg caacatttat acccagccgg
     1981 tggcgcagtg gcatgatcgc ggcgaacagg aagtgtttga atattgcctg gaagatggca
     2041 gcctgattcg cgcgaccaaa gatcataaat ttatgaccgt ggatggccag atgctgccga
     2101 ttgatgaaat ttttgaacgc gaactggatc tgatgcgcgt ggataacctg ccgaacggat
     2161 cagccggttc agcggccggt tcgggagtga aagtgattgg ccgccgcagc ctgggcgtgc
     2221 agcgcatttt tgatattggc ctgcgccagg atcataactt tctgctggcg aacggcgcga
     2281 ttgcggcgaa ctgctggaat ggcgccctga agggcgagat caagcagagg ctgaagctga
     2341 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     2401 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     2461 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2521 agctgtacaa gcatcatcac catcaccact aataatacta gtagcggccg ctgcaggagt
     2581 cactaagggt tagttagtta gattagcaga aagtcaaaag cctccgaccg gaggcttttg
     2641 actaaaactt cccttggggt tatcattggg gctcactcaa aggcggtaat cagataaaaa
     2701 aaatccttag ctttcgctaa ggatgatttc tgctagagat ggaatagact ggatggaggc
     2761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3121 ctgagcgtca gaccccttaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat
     3181 ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt tttcgaaggt tctctgagct
     3241 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     3301 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     3361 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     3421 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     3481 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     3541 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     3601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     3661 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     3721 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     3781 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     3841 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     3901 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     3961 agcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc
     4021 catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc
     4081 agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg
     4141 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
     4201 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     4261 ctgcttacat aaacagtaat acaaggggtg tttactagag gagattctca tgtttgacag
     4321 cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta acgcagtcag
     4381 gcaccgtgta tgaaatctaa caatgcgctc atcgtcattc tcggcaccgt caccctggac
     4441 gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga tatcgtccat
     4501 tccgacagta ttgccagtca ctatggcgtg ctgcttgcgc tctatgcgtt gatgcaattt
     4561 ctttgcgcac ccgttctcgg agccctgtcc gaccgctttg gccgccgtcc agtcctgctc
     4621 gcttcgctcc ttggagccac tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg
     4681 attctctacg ccggacgcat cgtggcgggc atcacgggtg ccacaggtgc ggttgctggt
     4741 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     4801 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggtgccatc
     4861 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct cctcctgggc
     4921 tgcttcctta tgcaggaatc gcataaggga gagcgccgtc cgatgccctt gcgtgccttc
     4981 aatccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     5041 gttttcttta tcatgcaact cgtaggacag gttccggcag cgctctgggt cattttcggc
     5101 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     5161 ttgcacgccc tcgctcaagc cttcgtcacg ggccccgcca ccaaacgttt cggcgagaag
     5221 caggccatta tcgcgggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg
     5281 acgcgcggct ggatggcctt ccccattatg attcttctcg cttccggcgg catcggtatg
     5341 cccgcgttgc aggccatgct gtcccgccaa gtagatgacg accatcaggg acagcttcaa
     5401 gggtcgctcg cggctcttac cagcctcact tcgatcattg gaccgctgat cgtcacggcg
     5461 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg tgccgccctt
     5521 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctaa
     5581 taatactagc tccggcaaaa aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa
     5641 accgaaaaga ttacttcgcg tt
//
