LOCUS       sequence-273521-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the SspDnaB M86 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  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 5683)
  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.R265
FEATURES             Location/Qualifiers
     source          1..5683
                     /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             2553..2570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2577..2597
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2633..2664
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3175..3719
                     /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     3620..3639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3836..3853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4335..4340,4341..4357,4358..4363)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4411..5601
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYVASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(4763..4782)
                     /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 cgtggatgct tgcatctcgg
     1861 gagatagttt gatcagcttg gcgagcacag ggaaaagagt tcctattaag gatttgttag
     1921 gcgaaaaaga ttttgaaata tgggcaatta atgaacagac gatgaagctg gaatcagcta
     1981 aagttagtcg tgtattttgt accggcaaaa agctagtcta tactctaaaa actcgactag
     2041 gtagaactat caaggcaaca gcaaatcata gatttttaac tattgatggt tggaaaagat
     2101 tagatgagct atctttaaaa gagcatattg ctctaccccg taaactagaa agctcctctt
     2161 tacaattggc accagaaata gaaaagttgc ctcagagtga tatttactgg gaccccatcg
     2221 tttctattac ggagactgga gtcgaagagg tttttgattt gactgtgcca ggactacgta
     2281 actttgtcgc caatgacatc attgtacata acagcgatct gggcgccctg aagggcgaga
     2341 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2401 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2461 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2521 actccaccgg cggcatggac gagctgtaca agcatcatca ccatcaccac taataatact
     2581 agtagcggcc gctgcaggag tcactaaggg ttagttagtt agattagcag aaagtcaaaa
     2641 gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca
     2701 aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagaga
     2761 tggaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2821 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2881 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2941 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3001 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3061 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3121 ttaacgtgag ttttcgttcc actgagcgtc agacccctta ataagatgat cttcttgaga
     3181 tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     3241 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     3301 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     3361 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     3481 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     3541 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     3601 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3661 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     3721 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     3781 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     3841 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     3901 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     3961 taagccagta tacactccgc tagcgctgag gtctgcctcg tgaagaaggt gttgctgact
     4021 cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga
     4081 gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt
     4141 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc
     4201 aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4261 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gtttactaga
     4321 ggagattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag
     4381 ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatt
     4441 ctcggcaccg tcaccctgga cgctgtaggc ataggcttgg ttatgccggt actgccgggc
     4501 ctcttgcggg atatcgtcca ttccgacagt attgccagtc actatggcgt gctgcttgcg
     4561 ctctatgcgt tgatgcaatt tctttgcgca cccgttctcg gagccctgtc cgaccgcttt
     4621 ggccgccgtc cagtcctgct cgcttcgctc cttggagcca ctatcgacta cgcgatcatg
     4681 gcgaccacac ccgtcctgtg gattctctac gccggacgca tcgtggcggg catcacgggt
     4741 gccacaggtg cggttgctgg tgcctatatc gccgacatca ccgatgggga agatcgggct
     4801 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     4861 gggggactgt tgggtgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
     4921 ggcctcaacc tcctcctggg ctgcttcctt atgcaggaat cgcataaggg agagcgccgt
     4981 ccgatgccct tgcgtgcctt caatccagtc agctccttcc ggtgggcgcg gggcatgact
     5041 atcgtcgccg cacttatgac tgttttcttt atcatgcaac tcgtaggaca ggttccggca
     5101 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     5161 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac gggccccgcc
     5221 accaaacgtt tcggcgagaa gcaggccatt atcgcgggca tggcggccga cgcgctgggc
     5281 tacgtcttgc tggcgttcgc gacgcgcggc tggatggcct tccccattat gattcttctc
     5341 gcttccggcg gcatcggtat gcccgcgttg caggccatgc tgtcccgcca agtagatgac
     5401 gaccatcagg gacagcttca agggtcgctc gcggctctta ccagcctcac ttcgatcatt
     5461 ggaccgctga tcgtcacggc gatttatgtc gcctcggcga gcacatggaa cgggttggca
     5521 tggattgtag gtgccgccct ttaccttgtc tgcctccccg cgttgcgtcg cggtgcatgg
     5581 agccgggcca cctcgaccta ataatactag ctccggcaaa aaaacgggca aggtgtcacc
     5641 accctgccct ttttctttaa aaccgaaaag attacttcgc gtt
//
