LOCUS       sequence-273512-        5725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the LLP-Pol 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  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 5725)
  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.R366
FEATURES             Location/Qualifiers
     source          1..5725
                     /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             2595..2612
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2619..2639
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2675..2706
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3217..3761
                     /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     3662..3681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3878..3895
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4377..4382,4383..4399,4400..4405)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4453..5643
                     /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(4805..4824)
                     /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 cgccaaggca tgcttttcag
     1861 gagataccga aatcttgacg ccatacggtt gggtgaagtt tgaagattac gacgaaagca
     1921 tgatgtgtgc tcaatacgat gaggttaccg gcaaaattag ttttacctat ccgaatgaaa
     1981 caatacacct gaaggatcag gacatctggg tgtatgagga tacaaatacc tccattcatg
     2041 cgaccggtaa ccacgatata ctgattcaga aaccaaacgg agatattgct aaggaaaagt
     2101 tttctaactt acaactgtta cagaaaggcg ataaacatag attcattaat gcgggatacg
     2161 tggattcggc cgaagaggta gacactctca tgcagcgtct ggttgggagc gcgggcagcg
     2221 cggcaggctc cggatctttc cctgggaagg cattccctac aggcccgagt tataaagggg
     2281 acgtgtattg cgtcaacgtc cccactcata atattgtaat tcgtcacaac gataaggtga
     2341 gcatacaggg caactgcaat ttcggcgccc tgaagggcga gatcaagcag aggctgaagc
     2401 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2461 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2521 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2581 acgagctgta caagcatcat caccatcacc actaataata ctagtagcgg ccgctgcagg
     2641 agtcactaag ggttagttag ttagattagc agaaagtcaa aagcctccga ccggaggctt
     2701 ttgactaaaa cttcccttgg ggttatcatt ggggctcact caaaggcggt aatcagataa
     2761 aaaaaatcct tagctttcgc taaggatgat ttctgctaga gatggaatag actggatgga
     2821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3181 ccactgagcg tcagacccct taataagatg atcttcttga gatcgttttg gtctgcgcgt
     3241 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggttttcgaa ggttctctga
     3301 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     3361 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     3421 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     3481 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     3541 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     3601 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     3661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     3721 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     3781 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     3841 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     3901 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     3961 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     4021 gctagcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag gcctgaatcg
     4081 ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg ttgtaggtgg
     4141 accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg tcgggaagat
     4201 gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt
     4261 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     4321 tgtctgctta cataaacagt aatacaaggg gtgtttacta gaggagattc tcatgtttga
     4381 cagcttatca tcgataagct ttaatgcggt agtttatcac agttaaattg ctaacgcagt
     4441 caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca ttctcggcac cgtcaccctg
     4501 gacgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc
     4561 cattccgaca gtattgccag tcactatggc gtgctgcttg cgctctatgc gttgatgcaa
     4621 tttctttgcg cacccgttct cggagccctg tccgaccgct ttggccgccg tccagtcctg
     4681 ctcgcttcgc tccttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg
     4741 tggattctct acgccggacg catcgtggcg ggcatcacgg gtgccacagg tgcggttgct
     4801 ggtgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     4861 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggtgcc
     4921 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctcctcctg
     4981 ggctgcttcc ttatgcagga atcgcataag ggagagcgcc gtccgatgcc cttgcgtgcc
     5041 ttcaatccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     5101 actgttttct ttatcatgca actcgtagga caggttccgg cagcgctctg ggtcattttc
     5161 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     5221 atcttgcacg ccctcgctca agccttcgtc acgggccccg ccaccaaacg tttcggcgag
     5281 aagcaggcca ttatcgcggg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     5341 gcgacgcgcg gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggt
     5401 atgcccgcgt tgcaggccat gctgtcccgc caagtagatg acgaccatca gggacagctt
     5461 caagggtcgc tcgcggctct taccagcctc acttcgatca ttggaccgct gatcgtcacg
     5521 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggtgccgcc
     5581 ctttaccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     5641 taataatact agctccggca aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt
     5701 aaaaccgaaa agattacttc gcgtt
//
