LOCUS       Exported                3538 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Red 
            Fluorescent Protein1 (mRFP1).
ACCESSION   .
VERSION     .
KEYWORDS    pNCS mRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3538)
  AUTHORS   Campbell RE, Tour O, Palmer AE, Steinbach PA, Baird GS, Zacharias 
            DA, Tsien RY
  TITLE     A monomeric red fluorescent protein.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7877-82.
  PUBMED    12060735
REFERENCE   2  (bases 1 to 3538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3538
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     57..76
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             107..129
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..166
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             170..202
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             206..229
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             236..913
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(366..384)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     634..653
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     828..851
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(976..994)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      990..1037
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1134..1589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1221..1240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1431..1452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1615..1719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1720..2580
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1938..1957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2751..3339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3240..3259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3493..3510
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat ctcgatcccg cgaaattaat
       61 acgactcact atagggagac cacaacggtt tccctctaga aataattttg tttaacttta
      121 agaaggagat atacatatgc ggggttctca tcatcatcat catcatggta tggctagcat
      181 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atccgatggc
      241 ctcctccgag gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt
      301 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      361 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
      421 ccctcagttc cagtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
      481 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
      541 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
      601 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
      661 ctgggaggcc tccaccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
      721 gatgaggctg aagctgaagg acggcggcca ctacgacgcc gaggtcaaga ccacctacat
      781 ggccaagaag cccgtgcagc tgcccggcgc ctacaagacc gacatcaagc tggacatcac
      841 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgcgccgagg gccgccactc
      901 caccggcgcc taagaattcg aagcttgatc cggctgctaa caaagcccga aaggaagctg
      961 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1021 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga
     1081 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1321 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1441 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1561 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2401 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3421 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3481 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaac
//
