LOCUS       sequence-296592-        5779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the TvoVMA 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5779)
  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 5779)
  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.R274
FEATURES             Location/Qualifiers
     source          1..5779
                     /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             2649..2666
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2673..2693
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2729..2760
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3271..3815
                     /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     3716..3735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3932..3949
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4431..4436,4437..4453,4454..4459)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4507..5697
                     /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(4859..4878)
                     /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 cgggagcaaa tgcgtatcag
     1861 gtgaaacacc agtttacctt gcggatggca agacaataaa aataaaggat ctatacagtt
     1921 ctgagagaaa aaaagaagat aacattgttg aggctggttc gggagaagag ataatacatc
     1981 taaaagatcc cattcaaata tattcttatg tggacgggac catagtcagg agcagatcaa
     2041 gacttctata caagggcaag agctcttatc ttgtgaggat agaaactatt ggcggaagat
     2101 cggtaagcgt tacaccagtt cacaaactct ttgtccttac ggaaaagggt atcgaagagg
     2161 ttatggcctc caacctaaag gtaggcgaca tgattgctgc tgtagcagaa agcgaatctg
     2221 aagcaagaga ctgcggaatg agcgaggaat gcgtgatgga agcagaagtt tatacgtcac
     2281 ttgaagcgac attcgataga gtaaagtcta tagcgtacga gaagggtgat tttgatgtat
     2341 acgatctttc cgtaccagaa tacggcagga actttatagg cggagaagga cttctcgtac
     2401 ttcacaacac tgtcattggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2461 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2521 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2581 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2641 tgtacaagca tcatcaccat caccactaat aatactagta gcggccgctg caggagtcac
     2701 taagggttag ttagttagat tagcagaaag tcaaaagcct ccgaccggag gcttttgact
     2761 aaaacttccc ttggggttat cattggggct cactcaaagg cggtaatcag ataaaaaaaa
     2821 tccttagctt tcgctaagga tgatttctgc tagagatgga atagactgga tggaggcgga
     2881 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2941 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3001 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3061 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3241 agcgtcagac cccttaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
     3301 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt cgaaggttct ctgagctacc
     3361 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     3421 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     3481 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     3541 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     3601 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     3661 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     3721 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     3781 cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac
     3841 ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat
     3901 ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata
     3961 tatcctgtat cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga
     4021 agcacttcac tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc
     4081 gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat
     4141 catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt
     4201 tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
     4261 tctgatcctt caactcagca aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa
     4321 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     4381 cttacataaa cagtaataca aggggtgttt actagaggag attctcatgt ttgacagctt
     4441 atcatcgata agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca
     4501 ccgtgtatga aatctaacaa tgcgctcatc gtcattctcg gcaccgtcac cctggacgct
     4561 gtaggcatag gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc
     4621 gacagtattg ccagtcacta tggcgtgctg cttgcgctct atgcgttgat gcaatttctt
     4681 tgcgcacccg ttctcggagc cctgtccgac cgctttggcc gccgtccagt cctgctcgct
     4741 tcgctccttg gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatt
     4801 ctctacgccg gacgcatcgt ggcgggcatc acgggtgcca caggtgcggt tgctggtgcc
     4861 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     4921 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg tgccatctcc
     4981 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctcct cctgggctgc
     5041 ttccttatgc aggaatcgca taagggagag cgccgtccga tgcccttgcg tgccttcaat
     5101 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtt
     5161 ttctttatca tgcaactcgt aggacaggtt ccggcagcgc tctgggtcat tttcggcgag
     5221 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     5281 cacgccctcg ctcaagcctt cgtcacgggc cccgccacca aacgtttcgg cgagaagcag
     5341 gccattatcg cgggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     5401 cgcggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cggtatgccc
     5461 gcgttgcagg ccatgctgtc ccgccaagta gatgacgacc atcagggaca gcttcaaggg
     5521 tcgctcgcgg ctcttaccag cctcacttcg atcattggac cgctgatcgt cacggcgatt
     5581 tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggtgc cgccctttac
     5641 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctaataa
     5701 tactagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc
     5761 gaaaagatta cttcgcgtt
//
