LOCUS       sequence-273505-        5695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the CP-Thy1 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  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 5695)
  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.R359
FEATURES             Location/Qualifiers
     source          1..5695
                     /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             2565..2582
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2589..2609
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2645..2676
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3187..3731
                     /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     3632..3651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3848..3865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4347..4352,4353..4369,4370..4375)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4423..5613
                     /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(4775..4794)
                     /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 caagcatcac tgttatagtt
     1861 ccgacactga ggtcctgaca agcgaaggct ggaaacgttg ggaagaagta agtatgaacg
     1921 attcgttcgc tactttaagc gcatccgggc aggtagagta ccagtaccct agcgaggtca
     1981 ttcaggaaga atatcagggg gacatggtaa gagctaaagc caagggcgta gaccttctgg
     2041 ttactccaaa ccacaaaatg ttggcctgta ttacaaccac ccgcgcagga cgtcggaaag
     2101 aaaatttttc gctgataccg gcggatcaat tgattggcgt gagccatgcg tataaaaagg
     2161 acggggactg gcgggctggt tctgctggtt ccgccgctgg aagcggagga gcttcaaaga
     2221 ccagttacgg ttgggagaaa tattcaggaa ccgtgtattg cgcgacggta ccgaatcata
     2281 ccctgtatgt acgtaggaat ggtaagccag tatggtcggg taactcagtc cttggcgccc
     2341 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     2401 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     2461 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2521 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagcatcat caccatcacc
     2581 actaataata ctagtagcgg ccgctgcagg agtcactaag ggttagttag ttagattagc
     2641 agaaagtcaa aagcctccga ccggaggctt ttgactaaaa cttcccttgg ggttatcatt
     2701 ggggctcact caaaggcggt aatcagataa aaaaaatcct tagctttcgc taaggatgat
     2761 ttctgctaga gatggaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2821 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2881 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2941 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3001 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3061 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagacccct taataagatg
     3181 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     3241 cttgcagggc ggttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     3301 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     3361 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     3421 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     3481 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     3541 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     3601 gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg
     3661 tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga
     3721 gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc
     3781 atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac
     3841 cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg
     3901 acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca
     3961 gtgccaacat agtaagccag tatacactcc gctagcgctg aggtctgcct cgtgaagaag
     4021 gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa gtgagggagc
     4081 cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac ttttgctttg
     4141 ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag
     4201 ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     4261 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     4321 gtgtttacta gaggagattc tcatgtttga cagcttatca tcgataagct ttaatgcggt
     4381 agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg
     4441 ctcatcgtca ttctcggcac cgtcaccctg gacgctgtag gcataggctt ggttatgccg
     4501 gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gtattgccag tcactatggc
     4561 gtgctgcttg cgctctatgc gttgatgcaa tttctttgcg cacccgttct cggagccctg
     4621 tccgaccgct ttggccgccg tccagtcctg ctcgcttcgc tccttggagc cactatcgac
     4681 tacgcgatca tggcgaccac acccgtcctg tggattctct acgccggacg catcgtggcg
     4741 ggcatcacgg gtgccacagg tgcggttgct ggtgcctata tcgccgacat caccgatggg
     4801 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     4861 ggccccgtgg ccgggggact gttgggtgcc atctccttgc atgcaccatt ccttgcggcg
     4921 gcggtgctca acggcctcaa cctcctcctg ggctgcttcc ttatgcagga atcgcataag
     4981 ggagagcgcc gtccgatgcc cttgcgtgcc ttcaatccag tcagctcctt ccggtgggcg
     5041 cggggcatga ctatcgtcgc cgcacttatg actgttttct ttatcatgca actcgtagga
     5101 caggttccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     5161 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     5221 acgggccccg ccaccaaacg tttcggcgag aagcaggcca ttatcgcggg catggcggcc
     5281 gacgcgctgg gctacgtctt gctggcgttc gcgacgcgcg gctggatggc cttccccatt
     5341 atgattcttc tcgcttccgg cggcatcggt atgcccgcgt tgcaggccat gctgtcccgc
     5401 caagtagatg acgaccatca gggacagctt caagggtcgc tcgcggctct taccagcctc
     5461 acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg
     5521 aacgggttgg catggattgt aggtgccgcc ctttaccttg tctgcctccc cgcgttgcgt
     5581 cgcggtgcat ggagccgggc cacctcgacc taataatact agctccggca aaaaaacggg
     5641 caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgtt
//
