LOCUS       Exported                5656 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses the split mCherry reporter interrupted by the NrdJ-1 
            intein (in cis).
ACCESSION   .
VERSION     .
KEYWORDS    pFP.R269
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5656)
  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 5656)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5656
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      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             2526..2543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2550..2570
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2606..2637
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3148..3692
                     /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     3593..3612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3809..3826
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4308..4336
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4384..5574
                     /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(4736..4755)
                     /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 caatccgtgt tgtctggttg
     1861 gtagcagcga aatcattacc cgtaattatg gtaaaaccac catcaaagaa gtggtcgaga
     1921 tcttcgataa cgacaaaaac attcaggtgc tggcctttaa tacccatacc gataatattg
     1981 aatgggcacc gattaaagca gcacagctga cccgtccgaa tgcagaactg gttgaactgg
     2041 aaattgatac cctgcatggt gttaaaacca ttcgttgtac accggatcat cctgtgtata
     2101 ccaaaaatcg tggttatgtt cgtgcagatg aactgaccga tgatgatgaa ctggtggttg
     2161 caattatgga agccaaaacc tatatcggca aactgaaaag ccgtaaaatt gtgagcaacg
     2221 aggataccta tgatattcag accagcaccc ataacttttt cgccaatgat attctggtgc
     2281 ataattcaga gatcggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2341 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2401 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2461 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2521 acaagcatca tcaccatcac cactaataat actagtagcg gccgctgcag gagtcactaa
     2581 gggttagtta gttagattag cagaaagtca aaagcctccg accggaggct tttgactaaa
     2641 acttcccttg gggttatcat tggggctcac tcaaaggcgg taatcagata aaaaaaatcc
     2701 ttagctttcg ctaaggatga tttctgctag agatggaata gactggatgg aggcggataa
     2761 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2821 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2881 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2941 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3001 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3061 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3121 gtcagacccc ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
     3181 ctctgaaaac gaaaaaaccg ccttgcaggg cggttttcga aggttctctg agctaccaac
     3241 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     3301 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     3361 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     3421 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     3481 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     3541 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     3601 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     3661 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     3721 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     3781 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     3841 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     3901 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     3961 gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat
     4021 ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg
     4081 tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct
     4141 gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc
     4201 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     4261 acataaacag taatacaagg ggtgtttact agaggagatt ctcatgtttg acagcttatc
     4321 atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     4381 tgtatgaaat ctaacaatgc gctcatcgtc attctcggca ccgtcaccct ggacgctgta
     4441 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac
     4501 agtattgcca gtcactatgg cgtgctgctt gcgctctatg cgttgatgca atttctttgc
     4561 gcacccgttc tcggagccct gtccgaccgc tttggccgcc gtccagtcct gctcgcttcg
     4621 ctccttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggattctc
     4681 tacgccggac gcatcgtggc gggcatcacg ggtgccacag gtgcggttgc tggtgcctat
     4741 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     4801 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggtgc catctccttg
     4861 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctcctcct gggctgcttc
     4921 cttatgcagg aatcgcataa gggagagcgc cgtccgatgc ccttgcgtgc cttcaatcca
     4981 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgttttc
     5041 tttatcatgc aactcgtagg acaggttccg gcagcgctct gggtcatttt cggcgaggac
     5101 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     5161 gccctcgctc aagccttcgt cacgggcccc gccaccaaac gtttcggcga gaagcaggcc
     5221 attatcgcgg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcgc
     5281 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg tatgcccgcg
     5341 ttgcaggcca tgctgtcccg ccaagtagat gacgaccatc agggacagct tcaagggtcg
     5401 ctcgcggctc ttaccagcct cacttcgatc attggaccgc tgatcgtcac ggcgatttat
     5461 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggtgccgc cctttacctt
     5521 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctaataatac
     5581 tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     5641 aagattactt cgcgtt
//
