LOCUS       sequence-273498-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the NrdA-2 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  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.R375
FEATURES             Location/Qualifiers
     source          1..5668
                     /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             2538..2555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2562..2582
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2618..2649
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3160..3704
                     /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     3605..3624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3821..3838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4320..4325,4326..4342,4343..4348)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4396..5586
                     /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(4748..4767)
                     /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 ctcaaacctt tgtctcacgg
     1861 gcgatgctaa aatcgacgta cttatcgata atattcctat ctcccagata tccctggagg
     1921 aagttgttaa cctgtttaat gaaggcaaag agatatatgt tttgtcttat aatattgata
     1981 ccaaggaggt tgaatataaa gaaatttctg acgcaggtct catcagtgaa tctgccgaag
     2041 tgctggaaat cattgacgaa gaaactgggc agaaaattgt ttgtacccct gatcataaag
     2101 tgtatactct gaaccggggg tacgtttctg ctaaggatct caaagaagac gatgagctgg
     2161 tgtttagcgg atcagccggc agcgcagctg gctcgggcgg tttaaaaatt ataaaacgtg
     2221 aatcaaagga accagtgttt gacattactg tcaaagataa tagcaatttt tttgccaata
     2281 atatcttggt gcataactgc aatgagggcg ccctgaaggg cgagatcaag cagaggctga
     2341 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2401 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     2461 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2521 tggacgagct gtacaagcat catcaccatc accactaata atactagtag cggccgctgc
     2581 aggagtcact aagggttagt tagttagatt agcagaaagt caaaagcctc cgaccggagg
     2641 cttttgacta aaacttccct tggggttatc attggggctc actcaaaggc ggtaatcaga
     2701 taaaaaaaat ccttagcttt cgctaaggat gatttctgct agagatggaa tagactggat
     2761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2821 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3001 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3121 gttccactga gcgtcagacc ccttaataag atgatcttct tgagatcgtt ttggtctgcg
     3181 cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttc gaaggttctc
     3241 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     3301 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     3361 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     3421 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     3481 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     3541 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     3601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     3661 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     3721 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     3781 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     3841 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
     3901 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
     3961 tccgctagcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa
     4021 tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg
     4081 tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa
     4141 gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg
     4201 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     4261 aactgtctgc ttacataaac agtaatacaa ggggtgttta ctagaggaga ttctcatgtt
     4321 tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa ttgctaacgc
     4381 agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcattctcgg caccgtcacc
     4441 ctggacgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc
     4501 gtccattccg acagtattgc cagtcactat ggcgtgctgc ttgcgctcta tgcgttgatg
     4561 caatttcttt gcgcacccgt tctcggagcc ctgtccgacc gctttggccg ccgtccagtc
     4621 ctgctcgctt cgctccttgg agccactatc gactacgcga tcatggcgac cacacccgtc
     4681 ctgtggattc tctacgccgg acgcatcgtg gcgggcatca cgggtgccac aggtgcggtt
     4741 gctggtgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4801 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggt
     4861 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctcctc
     4921 ctgggctgct tccttatgca ggaatcgcat aagggagagc gccgtccgat gcccttgcgt
     4981 gccttcaatc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     5041 atgactgttt tctttatcat gcaactcgta ggacaggttc cggcagcgct ctgggtcatt
     5101 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     5161 ggaatcttgc acgccctcgc tcaagccttc gtcacgggcc ccgccaccaa acgtttcggc
     5221 gagaagcagg ccattatcgc gggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     5281 ttcgcgacgc gcggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     5341 ggtatgcccg cgttgcaggc catgctgtcc cgccaagtag atgacgacca tcagggacag
     5401 cttcaagggt cgctcgcggc tcttaccagc ctcacttcga tcattggacc gctgatcgtc
     5461 acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggtgcc
     5521 gccctttacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     5581 acctaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct gccctttttc
     5641 tttaaaaccg aaaagattac ttcgcgtt
//
