LOCUS       sequence-158796-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dendra2 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  AUTHORS   Mohr MA, Argast P, Pantazis P
  TITLE     Labeling cellular structures in vivo using confined primed 
            conversion of photoconvertible fluorescent proteins.
  JOURNAL   Nat Protoc. 2016 Dec;11(12):2419-2431. doi: 10.1038/nprot.2016.134. 
            Epub 2016 Nov 3.
  PUBMED    27809312
REFERENCE   2  (bases 1 to 5756)
  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: 
            "10.1038/nprot.2016.134"; journalName: "Nat Protoc"; date: 
            "2016-12"; volume: "11"; issue: "12"; pages: "2419-2431"
COMMENT     Sequence Label: pGEX6P-1-Dendra2
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(184..189,190..205,206..212)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    220..236
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             259..912
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     870..892
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             919..942
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             952..1641
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     primer_bind     complement(1806..1828)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1973..1991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2059..2163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2164..2232,2233..3024)
                     /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(2382..2401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3195..3783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3684..3703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3937..3954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4027..4104
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4105..5187
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4124..4143)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5236..5241,5242..5259,5260..5266)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5274..5290
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5298..5314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5298..5314
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5310..5756,1..153)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLFDCTVHQCFWRQ
                     AAIGSCGMAVQVVNHCIIRVAQGALPFWIMFFAPTS"
     primer_bind     complement(5330..5347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5330..5346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5339..5361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttattcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
       61 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
      121 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
      181 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
      241 acacaggaaa cagtattcat gtcccctata ctaggttatt ggaaaattaa gggccttgtg
      301 caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag
      361 cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat
      421 cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat
      481 atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg
      541 cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac
      601 tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa
      661 gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg
      721 ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca
      781 aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa
      841 tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac
      901 catcctccaa aatcggatct ggaagttctg ttccaggggc ccctgggatc catgaacacc
      961 ccgggaatta acctgatcaa ggaggacatg cgcgtgaagg tgcacatgga gggcaacgtg
     1021 aacggccacg ccttcgtgat cgagggcgag ggcaagggca agccctacga gggcacccag
     1081 accgccaacc tgaccgtgaa ggagggcgcc cccctgccct tcagctacga catcctgacc
     1141 accgccgtgc actacggcaa ccgggtgttc accaagtacc ccgaggacat ccccgactac
     1201 ttcaagcaga gcttccccga gggctacagc tgggagcgca ccatgacctt cgaggacaag
     1261 ggcatctgca ccatccgcag cgacatcagc ctggagggcg actgcttctt ccagaacgtg
     1321 cgcttcaagg gcaccaactt cccccccaac ggccccgtga tgcagaagaa gaccctgaag
     1381 tgggagccca gcaccgagaa gctgcacgtg cgcgacggcc tgctggtggg caacatcaac
     1441 atggccctgc tgctggaggg cggcggccac tacctgtgcg acttcaagac cacctacaag
     1501 gccaagaagg tggtgcagct gcccgacgcc cacttcgtgg accaccgcat cgagatcctg
     1561 ggcaacgaca gcgactacaa caaggtgaag ctgtacgagc acgccgtggc ccgctacagc
     1621 cccctgccca gccaggtgtg gtccggcgac agcggcgtgt acaagtgaaa gcttaattag
     1681 ctgagcttgg actcctgttg aaagggcgaa ttctgcagat atccatcaca ctggcggccg
     1741 ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
     1801 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1861 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     1921 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
     1981 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2401 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2701 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3301 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3961 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4021 aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     4081 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     4141 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     4201 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     4261 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     4321 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     4381 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     4441 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     4501 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     4561 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     4621 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     4681 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     4741 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     4801 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     4861 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     4921 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     4981 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     5041 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     5101 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5161 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5221 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5281 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
     5341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5461 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
     5521 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
     5581 ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca
     5641 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
     5701 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt
//
