LOCUS       sequence-251416-        3550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of an EosFP variant, mEos2 (a photoconvertible 
            fluorescent protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3550)
  AUTHORS   McKinney SA, Murphy CS, Hazelwood KL, Davidson MW, Looger LL
  TITLE     A bright and photostable photoconvertible fluorescent protein.
  JOURNAL   Nat Methods. 2009 Feb . 6(2):131-3.
  PUBMED    19169260
REFERENCE   2  (bases 1 to 3550)
  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 
            Methods. 2009 Feb . 6(2):131-3."
COMMENT     Sequence Label: pRSETa mEos2
FEATURES             Location/Qualifiers
     source          1..3550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(650..668)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      664..711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      808..1263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(895..914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1105..1126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1289..1393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1394..1462,1463..2254)
                     /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(1612..1631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2425..3013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2914..2933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3167..3184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3269..3288
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3269..3287
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3319..3341
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3361..3378
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3382..3414
                     /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(3418..3426,3427..3441)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(3457..3550,1..587)
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
ORIGIN
        1 agccttttga gggaaaacag agtatggatc ttgaagtcaa agagggcgga cctctgcctt
       61 ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc gccaaatatc
      121 cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa
      181 gcttgacttt cgaagacggg ggcatttgca ttgccagaaa cgacataaca atggaagggg
      241 acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta
      301 tgcagaagaa gacgctgaaa tgggagccct ccactgagaa aatgtatgtg cgtgatggag
      361 tgctgacggg tgatattcat atggctttgt tgcttgaagg aaatgcccat taccgatgtg
      421 acttcagaac tacttacaaa gctaaggaga agggtgtcaa gttaccaggc taccactttg
      481 tggaccactg cattgagatt ttaagccatg acaaagatta caacaaggtt aagctgtatg
      541 agcatgctgt tgctcattct ggattgcctg acaatgccag acgataagaa ttcgaagctt
      601 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      661 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      721 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      781 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
      841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1261 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3241 ttaatgcagg atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
     3301 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct
     3361 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
     3421 ctgtacgacg atgacgataa ggatcgatgg ggatccatga gtgcgattaa gccagacatg
     3481 aagatcaaac tccgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat
     3541 ggtacaggca
//
