LOCUS       sequence-181645-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Plum 
            (mPlum).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  AUTHORS   Wang L, Jackson WC, Steinbach PA, Tsien RY
  TITLE     Evolution of new nonantibody proteins via iterative somatic 
            hypermutation.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Nov 30;101(48):16745-9. Epub 2004 Nov
            19.
  PUBMED    15556995
REFERENCE   2  (bases 1 to 5296)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2004-11-30"; volume: "101(48)"; pages: 
            "16745-9. Epub 2004 Nov 19"
COMMENT     Sequence Label: pBAD mPlum
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..303
                     /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     226..245
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             330..352
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             360..1037
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     primer_bind     761..780
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     955..978
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1097..1114)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1267..1353
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1445..1472
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1491..1582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1583..1651,1652..2443)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1801..1820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2485..2940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2572..2591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2782..2803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3051..3639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3540..3559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3793..3810
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3825..3965
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3891..3910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4051..4073
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4211..4230
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4410..5288)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
       61 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
      121 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
      181 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
      241 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
      301 catacccgtt tttttgggct agaaataatt ttgtttaact ttaagaagga gatatacata
      361 tggtgagcaa gggcgaggag gtcatcaagg agttcatgcg cttcaaggag cacatggagg
      421 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      481 gcacccagac cgccaggctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      541 tcctgtcccc tcagatcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      601 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
      661 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
      721 acaaggtgaa ggtgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
      781 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
      841 agatgaagat gaggctgagg ctgaaggacg gcggccacta cgacgccgag gtcaagacca
      901 cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg
      961 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc
     1021 gccactccac cggcgcccat catcatcatc atcagtaaaa gcttggctgt tttggcggat
     1081 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     1141 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     1201 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     1261 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1321 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     1381 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     1441 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     1501 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1561 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1621 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1681 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1741 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1801 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     1861 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1921 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1981 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2041 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2101 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     2161 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2221 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2281 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2341 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2401 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2461 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2521 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2581 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2641 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2701 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2761 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2821 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2881 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2941 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3181 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3661 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3721 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3781 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3841 ctccttacgc atctgtgcgg tatttcacac cgcatctggt gcactctcag tacaatctgc
     3901 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3961 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4021 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4081 cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc caacagtccc ccggccacgg
     4141 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     4201 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     4261 tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct tatcatcgat
     4321 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
     4381 aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact
     4441 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     4501 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     4561 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     4621 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     4681 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     4741 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     4801 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     4861 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     4921 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     4981 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     5041 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     5101 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     5161 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     5221 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     5281 tcagccatac ttttca
//
