LOCUS       sequence-273504-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Cro-RPB2 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  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 5689)
  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.R356
FEATURES             Location/Qualifiers
     source          1..5689
                     /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             2559..2576
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2583..2603
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2639..2670
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3181..3725
                     /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     3626..3645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3842..3859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4341..4346,4347..4363,4364..4369)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4417..5607
                     /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(4769..4788)
                     /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 cgataaattc tgccagaaat
     1861 atgatacact ggtacttact cacctgggat ggatcaaact tggcgaaatc gatataacta
     1921 ttcataaagt tgctacgttg gacaagcacg ataacatcat ctacgtttat cctacgtcta
     1981 aatttgaatt tgactatgac ggggactttt acgagcataa aaataatagc atcgatattg
     2041 aatgcacaat taatcataaa ttatactgca aatacaacct gagctcatca tttaccctga
     2101 ttccagcaga taaagtgtat ggtaacaaag tgattatgaa aaatatggaa aatgaagtga
     2161 tctggacgga tccgggttct gctggcagcg cagccgggtc ggggagtaaa gaacaaattc
     2221 acaactataa aggcaaagtg tattgtatag aagtgccaga ttcccatatt tattatatga
     2281 agaccagctc tatcactcca ccagtttgga taggcaattc aaccaaaggc gccctgaagg
     2341 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     2401 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     2461 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     2521 gccgccactc caccggcggc atggacgagc tgtacaagca tcatcaccat caccactaat
     2581 aatactagta gcggccgctg caggagtcac taagggttag ttagttagat tagcagaaag
     2641 tcaaaagcct ccgaccggag gcttttgact aaaacttccc ttggggttat cattggggct
     2701 cactcaaagg cggtaatcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgc
     2761 tagagatgga atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2821 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2881 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2941 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3001 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3061 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccttaataa gatgatcttc
     3181 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     3241 gggcggtttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     3301 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     3361 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     3481 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     3541 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     3601 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3661 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     3721 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     3781 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     3841 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     3901 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     3961 acatagtaag ccagtataca ctccgctagc gctgaggtct gcctcgtgaa gaaggtgttg
     4021 ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt
     4081 tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg
     4141 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat
     4201 ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
     4261 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgttt
     4321 actagaggag attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
     4381 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
     4441 gtcattctcg gcaccgtcac cctggacgct gtaggcatag gcttggttat gccggtactg
     4501 ccgggcctct tgcgggatat cgtccattcc gacagtattg ccagtcacta tggcgtgctg
     4561 cttgcgctct atgcgttgat gcaatttctt tgcgcacccg ttctcggagc cctgtccgac
     4621 cgctttggcc gccgtccagt cctgctcgct tcgctccttg gagccactat cgactacgcg
     4681 atcatggcga ccacacccgt cctgtggatt ctctacgccg gacgcatcgt ggcgggcatc
     4741 acgggtgcca caggtgcggt tgctggtgcc tatatcgccg acatcaccga tggggaagat
     4801 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     4861 gtggccgggg gactgttggg tgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     4921 ctcaacggcc tcaacctcct cctgggctgc ttccttatgc aggaatcgca taagggagag
     4981 cgccgtccga tgcccttgcg tgccttcaat ccagtcagct ccttccggtg ggcgcggggc
     5041 atgactatcg tcgccgcact tatgactgtt ttctttatca tgcaactcgt aggacaggtt
     5101 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     5161 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcacgggc
     5221 cccgccacca aacgtttcgg cgagaagcag gccattatcg cgggcatggc ggccgacgcg
     5281 ctgggctacg tcttgctggc gttcgcgacg cgcggctgga tggccttccc cattatgatt
     5341 cttctcgctt ccggcggcat cggtatgccc gcgttgcagg ccatgctgtc ccgccaagta
     5401 gatgacgacc atcagggaca gcttcaaggg tcgctcgcgg ctcttaccag cctcacttcg
     5461 atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg
     5521 ttggcatgga ttgtaggtgc cgccctttac cttgtctgcc tccccgcgtt gcgtcgcggt
     5581 gcatggagcc gggccacctc gacctaataa tactagctcc ggcaaaaaaa cgggcaaggt
     5641 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtt
//
