LOCUS       sequence-296583-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the split mCherry reporter interrupted by the SspDnaX 
            artificially mini-intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  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 5623)
  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.R271
FEATURES             Location/Qualifiers
     source          1..5623
                     /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             2493..2510
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2517..2537
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2573..2604
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3115..3659
                     /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     3560..3579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3776..3793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4275..4280,4281..4297,4298..4303)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4351..5541
                     /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(4703..4722)
                     /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 cattgacgaa tgcttaacgg
     1861 gggactcaca agttttgacc cgcaatggct taatgtccat tgacaatccc caaataaaag
     1921 ggcgagaagt tctgagctac aacgaaactc tacagcaatg ggaatataaa aaagttttaa
     1981 gatggcttga cagaggcgaa aagcaaacat tgtctattaa gacaaaaaat tctacagtac
     2041 ggtgtacggc taaccattta atcagaactg aacaaggatg gacgagagcg gaaaacatca
     2101 ctcccggaat gaagatacta tcccctgcgc cgcaatggca tacaaatttc gaggaagttg
     2161 agtccgtcac taagggtcaa gtggaaaaag tttatgacct ggaggtggaa gataatcaca
     2221 attttgttgc caatggctta ctagtccata actgtcacac tggcgccctg aagggcgaga
     2281 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2341 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2401 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2461 actccaccgg cggcatggac gagctgtaca agcatcatca ccatcaccac taataatact
     2521 agtagcggcc gctgcaggag tcactaaggg ttagttagtt agattagcag aaagtcaaaa
     2581 gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca
     2641 aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagaga
     2701 tggaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2761 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2821 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2881 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2941 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3061 ttaacgtgag ttttcgttcc actgagcgtc agacccctta ataagatgat cttcttgaga
     3121 tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     3181 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     3241 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     3301 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     3361 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     3421 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     3481 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     3541 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3601 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     3661 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     3721 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     3781 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     3841 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     3901 taagccagta tacactccgc tagcgctgag gtctgcctcg tgaagaaggt gttgctgact
     3961 cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga
     4021 gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt
     4081 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc
     4141 aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4201 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gtttactaga
     4261 ggagattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag
     4321 ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatt
     4381 ctcggcaccg tcaccctgga cgctgtaggc ataggcttgg ttatgccggt actgccgggc
     4441 ctcttgcggg atatcgtcca ttccgacagt attgccagtc actatggcgt gctgcttgcg
     4501 ctctatgcgt tgatgcaatt tctttgcgca cccgttctcg gagccctgtc cgaccgcttt
     4561 ggccgccgtc cagtcctgct cgcttcgctc cttggagcca ctatcgacta cgcgatcatg
     4621 gcgaccacac ccgtcctgtg gattctctac gccggacgca tcgtggcggg catcacgggt
     4681 gccacaggtg cggttgctgg tgcctatatc gccgacatca ccgatgggga agatcgggct
     4741 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     4801 gggggactgt tgggtgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
     4861 ggcctcaacc tcctcctggg ctgcttcctt atgcaggaat cgcataaggg agagcgccgt
     4921 ccgatgccct tgcgtgcctt caatccagtc agctccttcc ggtgggcgcg gggcatgact
     4981 atcgtcgccg cacttatgac tgttttcttt atcatgcaac tcgtaggaca ggttccggca
     5041 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     5101 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac gggccccgcc
     5161 accaaacgtt tcggcgagaa gcaggccatt atcgcgggca tggcggccga cgcgctgggc
     5221 tacgtcttgc tggcgttcgc gacgcgcggc tggatggcct tccccattat gattcttctc
     5281 gcttccggcg gcatcggtat gcccgcgttg caggccatgc tgtcccgcca agtagatgac
     5341 gaccatcagg gacagcttca agggtcgctc gcggctctta ccagcctcac ttcgatcatt
     5401 ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca
     5461 tggattgtag gtgccgccct ttaccttgtc tgcctccccg cgttgcgtcg cggtgcatgg
     5521 agccgggcca cctcgaccta ataatactag ctccggcaaa aaaacgggca aggtgtcacc
     5581 accctgccct ttttctttaa aaccgaaaag attacttcgc gtt
//
