LOCUS       sequence-148211-        3579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rsGreen0.8 in E. coli strains..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3579)
  AUTHORS   Duwe S, De Zitter E, Gielen V, Moeyaert B, Vandenberg W, Grotjohann 
            T, Clays K, Jakobs S, Van Meervelt L, Dedecker P
  TITLE     Expression-Enhanced Fluorescent Proteins Based on Enhanced Green 
            Fluorescent Protein for Super-resolution Microscopy.
  JOURNAL   ACS Nano. 2015 Oct 27;9(10):9528-41. doi: 10.1021/acsnano.5b04129. 
            Epub 2015 Sep 9.
  PUBMED    26308583
REFERENCE   2  (bases 1 to 3579)
  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"; doi: 
            "10.1021/acsnano.5b04129"; journalName: "ACS Nano"; date: 
            "2015-10-27- 27"; volume: "9"; issue: "10"; pages: "9528-41"
COMMENT     Sequence Label: pRSETb-rsGreen0.8
FEATURES             Location/Qualifiers
     source          1..3579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             111..128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             132..164
                     /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(168..176,177..191)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             198..917
                     /codon_start=1
                     /product="reversibly switchable enhanced green fluorescent 
                     protein (Grotjohann et al., 2011)"
                     /label=rsEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVLCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GYYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNAYITADKQKNGIR
                     SNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQNKLSKDPNEKRDHMVLL
                     EFVTASGITLGMDELYK"
     primer_bind     complement(243..264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     851..872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(980..998)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      994..1041
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1138..1593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1225..1244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1435..1456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1619..1723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1724..1792,1793..2584)
                     /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(1942..1961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2755..3343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3244..3263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3497..3514
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttactt taagaaggag atatacatat gcggggttct catcatcatc
      121 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
      181 atgacgataa ggatccgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      241 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
      301 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      361 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgctgtgc ttcgcccgct
      421 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      481 aggagcgcac catcagcttc aaggacgacg gctattacaa gacccgcgcc gaggtgaagt
      541 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      601 gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacgcc tatatcacgg
      661 ccgacaagca gaagaacggc atcaggtcta acttcaagat ccgccacaac gtcgaggacg
      721 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      781 tgctgcccga caaccactac ctgagcaccc agaataagct gagcaaagac cccaacgaga
      841 agcgcgatca catggtcctg ctggagttcg tgaccgcctc cgggatcact ctcggcatgg
      901 acgagctgta caagtaagaa ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa
      961 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1021 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg
     1081 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     1141 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1201 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1261 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1321 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1381 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1441 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1501 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1561 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2881 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3421 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3481 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3541 accgcctctc cccgcgcgtt ggccgattca ttaatgcag
//
