LOCUS       sequence-273511-        5692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the SspGyrB 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5692)
  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 5692)
  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.R272
FEATURES             Location/Qualifiers
     source          1..5692
                     /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             2096..2107
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2562..2579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2586..2606
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2642..2673
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3184..3728
                     /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     3629..3648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3845..3862
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4344..4349,4350..4366,4367..4372)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4420..5610
                     /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(4772..4791)
                     /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 cgcaggtggt tgtttttctg
     1861 gagatacatt agtcgcttta actgatggtc gtagcgttag ctttgagcaa ttggttgaag
     1921 aagaaaaaca aggaaaacaa aacttttgtt ataccatccg ccatgatggt tctatagggg
     1981 ttgaaaaaat catcaatgcc cgcaaaacaa aaactaatgc gaaggtaatc aaggttacgt
     2041 tggacaatgg tgagtctatt atttgcaccc cggatcataa attcatgttg cgggatggga
     2101 gctacaaatg tgcgatggat ttaactctcg atgattcgtt aatgccgtta caccgaaaaa
     2161 tttcgactac ggaagattct ggtcatgcga tggaagcagt attaaattac aatcacagaa
     2221 ttgtaaatat tgaagctgtg tcagaaacaa tcgatgttta tgatattgag gttccccaca
     2281 cccacaattt tgctttggca agcggagtgt ttgtccataa cagtgcaaaa ggcgccctga
     2341 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2401 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2461 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2521 agggccgcca ctccaccggc ggcatggacg agctgtacaa gcatcatcac catcaccact
     2581 aataatacta gtagcggccg ctgcaggagt cactaagggt tagttagtta gattagcaga
     2641 aagtcaaaag cctccgaccg gaggcttttg actaaaactt cccttggggt tatcattggg
     2701 gctcactcaa aggcggtaat cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     2761 tgctagagat ggaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2821 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2881 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2941 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3001 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3061 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc
     3181 ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt
     3241 gcagggcggt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     3301 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     3361 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     3421 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3481 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     3541 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     3601 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     3661 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     3721 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     3781 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     3841 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     3901 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     3961 ccaacatagt aagccagtat acactccgct agcgctgagg tctgcctcgt gaagaaggtg
     4021 ttgctgactc ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac
     4081 ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca
     4141 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     4201 gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     4261 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     4321 tttactagag gagattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt
     4381 ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc
     4441 atcgtcattc tcggcaccgt caccctggac gctgtaggca taggcttggt tatgccggta
     4501 ctgccgggcc tcttgcggga tatcgtccat tccgacagta ttgccagtca ctatggcgtg
     4561 ctgcttgcgc tctatgcgtt gatgcaattt ctttgcgcac ccgttctcgg agccctgtcc
     4621 gaccgctttg gccgccgtcc agtcctgctc gcttcgctcc ttggagccac tatcgactac
     4681 gcgatcatgg cgaccacacc cgtcctgtgg attctctacg ccggacgcat cgtggcgggc
     4741 atcacgggtg ccacaggtgc ggttgctggt gcctatatcg ccgacatcac cgatggggaa
     4801 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     4861 cccgtggccg ggggactgtt gggtgccatc tccttgcatg caccattcct tgcggcggcg
     4921 gtgctcaacg gcctcaacct cctcctgggc tgcttcctta tgcaggaatc gcataaggga
     4981 gagcgccgtc cgatgccctt gcgtgccttc aatccagtca gctccttccg gtgggcgcgg
     5041 ggcatgacta tcgtcgccgc acttatgact gttttcttta tcatgcaact cgtaggacag
     5101 gttccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     5161 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcacg
     5221 ggccccgcca ccaaacgttt cggcgagaag caggccatta tcgcgggcat ggcggccgac
     5281 gcgctgggct acgtcttgct ggcgttcgcg acgcgcggct ggatggcctt ccccattatg
     5341 attcttctcg cttccggcgg catcggtatg cccgcgttgc aggccatgct gtcccgccaa
     5401 gtagatgacg accatcaggg acagcttcaa gggtcgctcg cggctcttac cagcctcact
     5461 tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
     5521 gggttggcat ggattgtagg tgccgccctt taccttgtct gcctccccgc gttgcgtcgc
     5581 ggtgcatgga gccgggccac ctcgacctaa taatactagc tccggcaaaa aaacgggcaa
     5641 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tt
//
