LOCUS       sequence-273492-        5713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the Ama-Ter 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5713)
  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 5713)
  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.R360
FEATURES             Location/Qualifiers
     source          1..5713
                     /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             2583..2600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2607..2627
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2663..2694
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3205..3749
                     /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     3650..3669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3866..3883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4365..4370,4371..4387,4388..4393)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4441..5631
                     /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(4793..4812)
                     /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 ccatggtaaa aatattgcac
     1861 atagcacccc tgtgctgact gcgaaccgtg ggtgggtgac acatggggat ctagttcctg
     1921 gagatcaggt tttccatccg agcggcaaac ccgtggacgt gctggcgctt tcggatgagg
     1981 ctgtagatga ctacgtcgta acttttacga atggggaaaa aatcagatgc catgctaatc
     2041 atgaatggac agtttattcc cgcgcggata aacaggaaaa gacggtcgaa acaaaatggt
     2101 ttcttgagaa tacgaaccgc ggaactccac gctccttaac cgcgggaaat cgttttcagt
     2161 ttcaggttcc aaagacaaac gccctggaag gatccgcggg ttctgcggcc ggttctggcg
     2221 agcgccgcgt ttccatagag aaagtagaat atcttcctaa cggggagaaa ggccattgca
     2281 ttcaggtgga tagcccagac ggtctgtatt tagtagggaa gaaacttgtt gccacccaca
     2341 actccgagct gggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2401 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2461 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2521 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2581 agcatcatca ccatcaccac taataatact agtagcggcc gctgcaggag tcactaaggg
     2641 ttagttagtt agattagcag aaagtcaaaa gcctccgacc ggaggctttt gactaaaact
     2701 tcccttgggg ttatcattgg ggctcactca aaggcggtaa tcagataaaa aaaatcctta
     2761 gctttcgcta aggatgattt ctgctagaga tggaatagac tggatggagg cggataaagt
     2821 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2881 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2941 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3001 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3181 agacccctta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     3241 tgaaaacgaa aaaaccgcct tgcagggcgg ttttcgaagg ttctctgagc taccaactct
     3301 ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag
     3361 ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc
     3421 cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc
     3481 gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta
     3541 cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac
     3601 cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct
     3661 ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat
     3721 gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct
     3781 gttaagtatc ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc
     3841 tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct
     3901 gtatcacata ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact
     3961 tcactgacac cctcatcagt gccaacatag taagccagta tacactccgc tagcgctgag
     4021 gtctgcctcg tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca
     4081 gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga
     4141 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat
     4201 ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct
     4261 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     4321 taaacagtaa tacaaggggt gtttactaga ggagattctc atgtttgaca gcttatcatc
     4381 gataagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca ggcaccgtgt
     4441 atgaaatcta acaatgcgct catcgtcatt ctcggcaccg tcaccctgga cgctgtaggc
     4501 ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca ttccgacagt
     4561 attgccagtc actatggcgt gctgcttgcg ctctatgcgt tgatgcaatt tctttgcgca
     4621 cccgttctcg gagccctgtc cgaccgcttt ggccgccgtc cagtcctgct cgcttcgctc
     4681 cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg gattctctac
     4741 gccggacgca tcgtggcggg catcacgggt gccacaggtg cggttgctgg tgcctatatc
     4801 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     4861 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggtgccat ctccttgcat
     4921 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tcctcctggg ctgcttcctt
     4981 atgcaggaat cgcataaggg agagcgccgt ccgatgccct tgcgtgcctt caatccagtc
     5041 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgttttcttt
     5101 atcatgcaac tcgtaggaca ggttccggca gcgctctggg tcattttcgg cgaggaccgc
     5161 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     5221 ctcgctcaag ccttcgtcac gggccccgcc accaaacgtt tcggcgagaa gcaggccatt
     5281 atcgcgggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgcggc
     5341 tggatggcct tccccattat gattcttctc gcttccggcg gcatcggtat gcccgcgttg
     5401 caggccatgc tgtcccgcca agtagatgac gaccatcagg gacagcttca agggtcgctc
     5461 gcggctctta ccagcctcac ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc
     5521 gcctcggcga gcacatggaa cgggttggca tggattgtag gtgccgccct ttaccttgtc
     5581 tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgaccta ataatactag
     5641 ctccggcaaa aaaacgggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
     5701 attacttcgc gtt
//
