LOCUS       sequence-208242-        3580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for the fluorescent protein enhanced YFP (eYFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3580)
  AUTHORS   Sarabipour S, King C, Hristova K
  TITLE     Uninduced high-yield bacterial expression of fluorescent proteins.
  JOURNAL   Anal Biochem. 2014 Mar 15;449:155-7. doi: 10.1016/j.ab.2013.12.027. 
            Epub 2013 Dec 28.
  PUBMED    24378720
REFERENCE   2  (bases 1 to 3580)
  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: "Anal 
            Biochem."; date: "2014-03-15"; volume: "449:155-7. doi"; pages: " 
            10.1016/j.ab.2013.12.027. Epub 2013 Dec 28"
COMMENT     Sequence Label: pRSET-B YFP
FEATURES             Location/Qualifiers
     source          1..3580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     854..873
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        854..872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             904..926
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             946..963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             967..999
                     /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             join(1003..1011,1012..1026)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1033..1035,1036..1038,1039..1752)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1078..1099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1339..1358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1686..1707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1815..1833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1829..1876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1973..2428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2060..2079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2270..2291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2454..2558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2559..2627,2628..3419)
                     /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(2777..2796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      781 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatctc
      841 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat
      901 aattttgttt aactttaaga aggagatata catatgcggg gttctcatca tcatcatcat
      961 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
     1021 gataaggatc caatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1081 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1141 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1201 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     1261 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1321 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1381 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1441 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1501 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1561 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1621 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     1681 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1741 ctgtacaagt aagaattcga agcttgatcc ggctgctaac aaagcccgaa aggaagctga
     1801 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1861 cttgaggggt tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag
     1921 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     1981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2341 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2401 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2461 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2521 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2581 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2641 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2701 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2761 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2821 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2881 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2941 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3001 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3061 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3121 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3181 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3241 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3301 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3361 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3421 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3481 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3541 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
//
