LOCUS       sequence-273517-        5737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Cro-Top2 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5737)
  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 5737)
  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.R357
FEATURES             Location/Qualifiers
     source          1..5737
                     /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             2607..2624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2631..2651
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2687..2718
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3229..3773
                     /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     3674..3693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3890..3907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4389..4394,4395..4411,4412..4417)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4465..5655
                     /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(4817..4836)
                     /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 cggcagtaag tgtatatcct
     1861 ccgataccaa cgtgctgata tggaacagta aaatctctaa aaaagccaaa gacattcaaa
     1921 tcggtgatat actggtaggt gacgatggta acaagcgcaa tgttatcgat gttagctttg
     1981 gaaaaggaca gatgtacaaa atcattcagt ccaaaggtga aaattatagc gttaataaca
     2041 accatactct tacgctgatg atgccactgc acaaagtaat tagcaatagt aacaataagc
     2101 tgaaattact ctggtgggat aatgtgtata aaattattaa atccaaaccg attgatgtgc
     2161 aagaaaacaa aattagcgat gatgaaattc tcctgaacag tgtctatcat gaccgcggct
     2221 cagccggtag tgccgctggt tccgggctgg attcgccagg gtatatcacg gttgagaaag
     2281 atggtattgg cgattatgtg agtattacag tggatacgac gacgaatcaa cgatttctga
     2341 ttaacgattt tacggtcacg cataattgct caaacggcgc cctgaagggc gagatcaagc
     2401 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2461 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2521 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2581 ccggcggcat ggacgagctg tacaagcatc atcaccatca ccactaataa tactagtagc
     2641 ggccgctgca ggagtcacta agggttagtt agttagatta gcagaaagtc aaaagcctcc
     2701 gaccggaggc ttttgactaa aacttccctt ggggttatca ttggggctca ctcaaaggcg
     2761 gtaatcagat aaaaaaaatc cttagctttc gctaaggatg atttctgcta gagatggaat
     2821 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2881 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2941 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3001 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3061 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3121 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3181 tgagttttcg ttccactgag cgtcagaccc cttaataaga tgatcttctt gagatcgttt
     3241 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggttttcg
     3301 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     3361 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     3421 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     3481 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     3541 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     3601 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     3661 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     3721 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     3781 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
     3841 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     3901 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
     3961 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
     4021 agtatacact ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc
     4081 aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt
     4141 tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt
     4201 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa
     4261 gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     4321 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgtttac tagaggagat
     4381 tctcatgttt gacagcttat catcgataag ctttaatgcg gtagtttatc acagttaaat
     4441 tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt cattctcggc
     4501 accgtcaccc tggacgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg
     4561 cgggatatcg tccattccga cagtattgcc agtcactatg gcgtgctgct tgcgctctat
     4621 gcgttgatgc aatttctttg cgcacccgtt ctcggagccc tgtccgaccg ctttggccgc
     4681 cgtccagtcc tgctcgcttc gctccttgga gccactatcg actacgcgat catggcgacc
     4741 acacccgtcc tgtggattct ctacgccgga cgcatcgtgg cgggcatcac gggtgccaca
     4801 ggtgcggttg ctggtgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     4861 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     4921 ctgttgggtg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     4981 aacctcctcc tgggctgctt ccttatgcag gaatcgcata agggagagcg ccgtccgatg
     5041 cccttgcgtg ccttcaatcc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     5101 gccgcactta tgactgtttt ctttatcatg caactcgtag gacaggttcc ggcagcgctc
     5161 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     5221 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcacgggccc cgccaccaaa
     5281 cgtttcggcg agaagcaggc cattatcgcg ggcatggcgg ccgacgcgct gggctacgtc
     5341 ttgctggcgt tcgcgacgcg cggctggatg gccttcccca ttatgattct tctcgcttcc
     5401 ggcggcatcg gtatgcccgc gttgcaggcc atgctgtccc gccaagtaga tgacgaccat
     5461 cagggacagc ttcaagggtc gctcgcggct cttaccagcc tcacttcgat cattggaccg
     5521 ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt
     5581 gtaggtgccg ccctttacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg
     5641 gccacctcga cctaataata ctagctccgg caaaaaaacg ggcaaggtgt caccaccctg
     5701 ccctttttct ttaaaaccga aaagattact tcgcgtt
//
