LOCUS       sequence-403151-        5758 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 5758)
  AUTHORS   Sugita K, Onishi I, Nakayama R, Ishibashi S, Ikeda M, Inoue M, 
            Narita R, Oshima S, Shimizu K, Saito S, Sato S, Moriarity BS, 
            Yamamoto K, Largaespada DA, Kitagawa M, Kurata M
  TITLE     Indirect CRISPR screening with photoconversion revealed key factors 
            of drug resistance with cell-cell interactions.
  JOURNAL   Commun Biol. 2023 Jun 1;6(1):582. doi: 10.1038/s42003-023-04941-9.
  PUBMED    37264057
REFERENCE   2  (bases 1 to 5758)
  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: "Commun 
            Biol."; date: "2023-06-1"; volume: "6(1):582. doi"; pages: " 
            10.1038/s42003-023-04941-9"
COMMENT     Sequence Label: pDendra2-Hygro
FEATURES             Location/Qualifiers
     source          1..5758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             628..1662
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1644..1652
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1663..1716
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1732..2424
                     /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"
     polyA_signal    2546..2667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2583..2602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2637..2656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2674..3129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3023..3042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3156..3260
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3262..3619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3262..3453
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3285..3305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3470..3605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3654..4448
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3654..4448
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3708..3727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4318..4337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4636..4655)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4680..4727
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5056..5644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5545..5564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatg aagaagcccg aactcaccgc taccagcgtt
      661 gaaaaatttc tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc
      721 gaagagagcc gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat
      781 agctgcgctg atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca
      841 ctacccatcc ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc
      901 agtagacgcg cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg
      961 ttacagcctg tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc
     1021 ggcttcggcc cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt
     1081 tgcgccattg ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc
     1141 gccagcgtag ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg
     1201 cgccacctcg tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc
     1261 accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac
     1321 atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc
     1381 cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg
     1441 gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc
     1501 cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc
     1561 cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc
     1621 cggcccagta cacgaccgcg cgctaaggag gtaggtcgag ttgagggcag aggaagtctg
     1681 ttaacatgcg gtgacgtcga ggagaatcct ggcccaccac cggtcgccac catgaacacc
     1741 ccgggaatta acctgatcaa ggaggacatg cgcgtgaagg tgcacatgga gggcaacgtg
     1801 aacggccacg ccttcgtgat cgagggcgag ggcaagggca agccctacga gggcacccag
     1861 accgccaacc tgaccgtgaa ggagggcgcc cccctgccct tcagctacga catcctgacc
     1921 accgccgtgc actacggcaa ccgggtgttc accaagtacc ccgaggacat ccccgactac
     1981 ttcaagcaga gcttccccga gggctacagc tgggagcgca ccatgacctt cgaggacaag
     2041 ggcatctgca ccatccgcag cgacatcagc ctggagggcg actgcttctt ccagaacgtg
     2101 cgcttcaagg gcaccaactt cccccccaac ggccccgtga tgcagaagaa gaccctgaag
     2161 tgggagccca gcaccgagaa gctgcacgtg cgcgacggcc tgctggtggg caacatcaac
     2221 atggccctgc tgctggaggg cggcggccac tacctgtgcg acttcaagac cacctacaag
     2281 gccaagaagg tggtgcagct gcccgacgcc cacttcgtgg accaccgcat cgagatcctg
     2341 ggcaacgaca gcgactacaa caaggtgaag ctgtacgagc acgccgtggc ccgctacagc
     2401 cccctgccca gccaggtgtg gtaagcggcc gcgactctag atcataatca gccataccac
     2461 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2521 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2641 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2701 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2761 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2821 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2881 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2941 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3001 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3061 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3121 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3241 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3301 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3481 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3541 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3601 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3661 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3721 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3781 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3841 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3901 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3961 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4021 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4081 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4141 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4201 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4261 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4321 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4381 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4441 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4621 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4681 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4741 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4801 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4861 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5161 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5701 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
