LOCUS       sequence-199008-        7023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of fluorescent protein Dendra2 used for 
            transfection efficiency control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7023)
  AUTHORS   Hsu AY, Wang D, Gurol T, Zhou W, Zhu X, Lu HY, Deng Q
  TITLE     Overexpression of microRNA-722 fine-tunes neutrophilic inflammation 
            through inhibiting Rac2 in zebrafish.
  JOURNAL   Dis Model Mech. 2017 Sep 27. pii: dmm.030791. doi: 
            10.1242/dmm.030791.
  PUBMED    28954734
REFERENCE   2  (bases 1 to 7023)
  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: "Dis Model 
            Mech. 2017 Sep 27. pii: dmm.030791. doi: 10.1242/dmm.030791."
COMMENT     Sequence Label: pcDNA3.1 Dendra2
FEATURES             Location/Qualifiers
     source          1..7023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      271..699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(358..377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     568..589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(708..728)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        713..1042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      893..1028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     955..974
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1109..1903
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1163..1182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1773..1792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2077..2198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2114..2133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2168..2187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2247..2263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2247..2263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2260..2282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2271..2287
                     /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)."
     promoter        complement(join(2295..2301,2302..2319,2320..2325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2340..2361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2478..2495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2649..3237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2729..2748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3408..4199,4200..4268))
                     /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     4031..4050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4269..4373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4448..4467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4639..5018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5019..5222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5173..5193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5267..5286
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5267..5285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5321..5330
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6294..6986
                     /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(join(7018..7023,1..12))
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
       61 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
      121 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
      181 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
      241 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
      301 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      361 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      421 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      481 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      541 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      601 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
      661 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
      721 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      781 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      841 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      901 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      961 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     1021 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     1081 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1141 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1201 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     1261 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     1321 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     1381 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     1441 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     1501 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     1561 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1621 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1681 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     1741 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1801 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1861 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1921 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     1981 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     2041 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     2101 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2161 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     2221 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2281 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2341 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2401 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2461 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2521 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2821 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3061 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3721 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3841 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4021 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4261 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4381 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     4441 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     4501 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     4561 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     4621 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     4681 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4741 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4801 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4861 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4921 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4981 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     5041 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5101 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5161 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5221 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     5281 atagggagac ccaagctggc tagcgtttaa acttaagctt gccaccatgg atgaggaaat
     5341 cgctgccctg gtcgttgaca acggctccgg tatgtgcaaa gccggttttg ctggagatga
     5401 tgcccctcgt gctgttttcc cctccattgt tggacgaccc agacatcagg tacatttaat
     5461 aaaccatcct aataacatgt taatcatatt gaaccacatt aaactctcat ttgtgcattt
     5521 tacatctttt aaaaccgatt tatattcgtt tttagataat gaagtttatt aaagtgttaa
     5581 ttcacagctt tattgcttgt tttgagaatc tgcttgcata agtgattttt aaacttgatc
     5641 tgattttcgt ttgtttcgaa gttcttcagc tttgaaagtc agatcttcaa ttcacagcca
     5701 tgccatagtt ttaggataca gtataagtag acgaattggg atgcatcgta gattttatgc
     5761 tgtgtttgtt atggtttttc agtggtagag attttcagag tattattagg gttatagtca
     5821 gtgtcaggtt tatttatcgt cagcgtgatg ggcaaaaaac agctggcagt acgggctatg
     5881 tcttcgtcta gaagaagaca aagccaactg ctgttactct cgcccagaaa tctgcgccaa
     5941 catggacaca cccagtggag caagccatct gattttcatg gtcagattgt aaagaagcaa
     6001 ttactcttgg acagtttaag cagaaaagcc accaagtcaa gtcactggtc atatgatgaa
     6061 ctcgaagatc taataaaagt taatttaagc gttcattaca ataaattcaa gtgaatctgt
     6121 tttaagctag cctttaacgt ttttaaattc cttcaaacaa attagttcct ctttgcagtt
     6181 cctcatttct cggtgagctg gtgctttaat ccctttactt tttcttcctc tagggagtga
     6241 tggttggcat gggacgaatt aattcctgca gcccgggggt accggtcgcc accatgaaca
     6301 ccccgggaat taacctgatc aaggaggaca tgcgcgtgaa ggtgcacatg gagggcaacg
     6361 tgaacggcca cgccttcgtg atcgagggcg agggcaaggg caagccctac gagggcaccc
     6421 agaccgccaa cctgaccgtg aaggagggcg cccccctgcc cttcagctac gacatcctga
     6481 ccaccgccgt gcactacggc aaccgggtgt tcaccaagta ccccgaggac atccccgact
     6541 acttcaagca gagcttcccc gagggctaca gctgggagcg caccatgacc ttcgaggaca
     6601 agggcatctg caccatccgc agcgacatca gcctggaggg cgactgcttc ttccagaacg
     6661 tgcgcttcaa gggcaccaac ttccccccca acggccccgt gatgcagaag aaaaccctga
     6721 agtgggagcc cagcaccgag aagctgcacg tgcgcgacgg cctgctggtg ggcaacatca
     6781 acatggccct gctgctggag ggcggcggcc actacctgtg cgacttcaag accacctaca
     6841 aggccaagaa ggtggtgcag ctgcccgacg cccacttcgt ggaccaccgc atcgagatcc
     6901 tgggcaacga cagcgactac aacaaggtga agctgtacga gcacgccgtg gcccgctaca
     6961 gccccctgcc cagccaggtg tggtaatcta gagggcccgt ttaaacccgc tgatcagcct
     7021 cga
//
