LOCUS       sequence-304854-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the SaP-dpol 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  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 5707)
  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.R373
FEATURES             Location/Qualifiers
     source          1..5707
                     /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             2577..2594
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2601..2621
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2657..2688
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3199..3743
                     /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     3644..3663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3860..3877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4359..4364,4365..4381,4382..4387)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4435..5625
                     /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(4787..4806)
                     /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 cggaaaggcc tgcctgcatc
     1861 gacacactca ggttcttact gatggcggat ttaaagatat catggccgtt accagtactg
     1921 ataaagtctg gagcggcgaa aaatgggtta atactaaggg ggcacatctg atggggtgga
     1981 aaccggttat taatgtcgat ggtgttctca tgaccgaaga tcacaaaatt ctcacccaca
     2041 gctggaaaca ggcaaagcag ctggtttcta acaaatatat gatggatcgt gctctcgaaa
     2101 taggcatgga tgcatggctc tcttgtgcga gttatcaaaa tgataaagcc aaggacaatt
     2161 acagtagtaa cgttattgtg gaacggtgcc tgggcggcag tgccgggtct gctgcgggct
     2221 caggcaatct gaacggcaaa ctgacgtact gtgaaccagt ttatgatcta atagatgtgg
     2281 aggacggtaa ccgattccta attgcgtcag attccggttt tttagtggct cataactgcg
     2341 aactgggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2401 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2461 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2521 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagcatc
     2581 atcaccatca ccactaataa tactagtagc ggccgctgca ggagtcacta agggttagtt
     2641 agttagatta gcagaaagtc aaaagcctcc gaccggaggc ttttgactaa aacttccctt
     2701 gtggttatca ttggggctca ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc
     2761 gctaaggatg atttctgcta gagatggaat agactggatg gaggcggata aagttgcagg
     2821 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2881 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2941 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3001 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3181 cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     3241 cgaaaaaacc gccttgcagg gcggttttcg aaggttctct gagctaccaa ctctttgaac
     3301 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     3361 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     3421 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3481 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     3541 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     3601 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     3661 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     3721 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     3781 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     3841 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     3901 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     3961 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc tgaggtctgc
     4021 ctcgtgaaga aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga
     4081 aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga
     4141 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
     4201 actcagcaaa agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt
     4261 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     4321 gtaatacaag gggtgtttac tagaggagat tctcatgttt gacagcttat catcgataag
     4381 ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     4441 tctaacaatg cgctcatcgt cattctcggc accgtcaccc tggacgctgt aggcataggc
     4501 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagtattgcc
     4561 agtcactatg gcgtgctgct tgcgctctat gcgttgatgc aatttctttg cgcacccgtt
     4621 ctcggagccc tgtccgaccg ctttggccgc cgtccagtcc tgctcgcttc gctccttgga
     4681 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggattct ctacgccgga
     4741 cgcatcgtgg cgggcatcac gggtgccaca ggtgcggttg ctggtgccta tatcgccgac
     4801 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     4861 ggtatggtgg caggccccgt ggccggggga ctgttgggtg ccatctcctt gcatgcacca
     4921 ttccttgcgg cggcggtgct caacggcctc aacctcctcc tgggctgctt ccttatgcag
     4981 gaatcgcata agggagagcg ccgtccgatg cccttgcgtg ccttcaatcc agtcagctcc
     5041 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtttt ctttatcatg
     5101 caactcgtag gacaggttcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     5161 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     5221 caagccttcg tcacgggccc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcg
     5281 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg cggctggatg
     5341 gccttcccca ttatgattct tctcgcttcc ggcggcatcg gtatgcccgc gttgcaggcc
     5401 atgctgtccc gccaagtaga tgacgaccat cagggacagc ttcaagggtc gctcgcggct
     5461 cttaccagcc tcacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     5521 gcgagcacat ggaacgggtt ggcatggatt gtaggtgccg ccctttacct tgtctgcctc
     5581 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctaataata ctagctccgg
     5641 caaaaaaacg ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact
     5701 tcgcgtt
//
