LOCUS       sequence-193379-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty vector for overexpression of microRNA and Dendra2 in 
            Zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7597)
  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 7597)
  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: Tol2-lyzC-Vector-Dendra2
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(561..572,573..710)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     regulatory      3139..3148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4111..4803
                     /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    4914..5035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4951..4970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5005..5024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(5159..5346,5347..5358)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5413..5433)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5413..5431)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5452..5468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5452..5468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5465..5487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5476..5492
                     /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(5500..5506,5507..5524,5525..5530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5545..5566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5683..5700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5854..6442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5934..5953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6613..7404,7405..7473))
                     /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     7236..7255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7474..7578)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggccac aatgacggct actacatggt
      361 gccattcctt cctctttata ggaatggaga ctacctcctg tccaacaagg ctcttggata
      421 cgagtacgcc tacctgttgg acccaggtca ttgcacaaca ccagaaatgc cctctgatct
      481 gcaaaagacg tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag
      541 gtttgtccag gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa
      601 agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct
      661 tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa
      721 aaggctagca agatctgctc gagagtttga cttgggttga ttatagcaaa tagtggaaca
      781 caaacagatt ttgaaatatt cttgcaaatt tatttattaa atccattcaa atattgtcat
      841 tttttgtact gtgtaattta tattgatttg ctcatacaat atttacattt cgcttaaaca
      901 atacactatt atttatattt aaaaatagct atatataaat caaatgtata gctctttttt
      961 acacaattaa tcaaagctaa aatatttgtc acacaaatac agtaagccac ttcacaatga
     1021 tatcttcaaa atctgtgctt gactgaatca tttcatttat tgttttgtaa ttcacattat
     1081 accttacgtt ctccaggcac agtttgaatt attgataaat gaagataata catctgattt
     1141 aattagtctc ctttgtttat aataacttta atctaatttg atagtaataa ttccagcaat
     1201 acatccatta caataaaaat cgctttttaa ttttgcacat tagtttgccg gtatttgttt
     1261 aaagtttcac catactttta aatcatgtgc tatttgaaaa gggctgttat gactggaatt
     1321 aaatgttttt ttttttatat cagtgtttta gattgaagta caatatttat aggtaaaatt
     1381 caccagcata tgcttaaaga tagccatgct ccaggtcctt gtgtccttct aaggtctgtt
     1441 tgtggtttaa gctgtttgtt gacacacgct tgtcatctcc atttttctgt gcagttattg
     1501 tcagtttatt gcacactggg ttgcattaag ctgcagtttt ttcacaacag ttttccacat
     1561 tgtaacagtc agtctccaaa ttattccatc agtcctcaaa tttagtcatt tgaggccatg
     1621 aaaacactta aaactacata aaggtagttt ggcattaggt tttagacttg ttcttgtttg
     1681 gcaatattgc aacctgtgta aaataatttg attctttttt tttttttttt ttttttaatt
     1741 ttcagaaaga aatggtccaa aaggagctat gtgaacattt atccatgtgt ttaatgtttt
     1801 tttttttatt attaaaccac ttctgactta acttttttta gttattctat tttgtgacga
     1861 gtctgaatgg ccaaagtact tttgaatcca aaatcttact agtaaaatgt taaatcttac
     1921 tagcagtgtt gggtataatt tgtaacaaag ttaaattcca tatctgctaa gtgaaaaagt
     1981 aaagtaagag attgctgtaa atatttaggt aatttgtttt attaatacaa tatagaaagt
     2041 aacaaaacac aatgtaaaaa aatcaataaa gaaagtaaat aataaaagta atgataaact
     2101 aatgatctaa acaaaattag taataaaatt tatacaattc agggagggta aagaatttca
     2161 cctaatagtc aaactctgta ttaaataata taaaatattt gttcttagtc tgaagaggag
     2221 actgtgagag aaagcttact ggttattaaa ttagaaaatt tgtaacttgc agattaaagc
     2281 tgtgtgcctt gcagattaaa ataatttaaa taatttaata aaggccatgt ttgagacaaa
     2341 tgtaaaaaca taataaatgt taagaatttc agatattgag attttacaat ttaaaaaaag
     2401 caataaagta atagcagaga cctacactct taaagtaaat tactcaacac tgcttactag
     2461 taacttttgg aatagttact agtatccatt gaacaaatac tagaatctaa ttaaattgtg
     2521 tgagtgtcca atgaaaagat tcttgtattt aaaacagtac cagtatctat atatgtacaa
     2581 aatttagtgc tagcaaaatg caaataaata ctactgcata taatggattg cattgtaaat
     2641 cgacttgcca taatacattt atttcctatt tttactttta acaaaaaata agttcactga
     2701 gtatttcatg tcattaaatc gcattaaacc tcttgcatgc tttatgtaca ctagtgcagt
     2761 tcgctaaatg accaccggtg tcactaataa cacgatcgtt tctgaattct gcattaaacg
     2821 ccatttatac gttgctctta ttagtaaaca atttctccat catgagctcg accagtattg
     2881 catatttcat cctcttaaat ccagagttgt ttcttttaat cactttctct tcatgataaa
     2941 agcagttttt ggttttcaga agtttacaac aaacccagtc agttattgcc tcagcaaaca
     3001 ctggagcaac tgcacacttg tgaaaagctt cttgcgacac aaacctttct ctggacttat
     3061 aaataaaggc attcagcagc tccaccatac acctccactg gtcttgataa agcagatatc
     3121 agcagtgata caggtaccgc caccatggat gaggaaatcg ctgccctggt cgttgacaac
     3181 ggctccggta tgtgcaaagc cggttttgct ggagatgatg cccctcgtgc tgttttcccc
     3241 tccattgttg gacgacccag acatcaggta catttaataa accatcctaa taacatgtta
     3301 atcatattga accacattaa actctcattt gtgcatttta catcttttaa aaccgattta
     3361 tattcgtttt tagataatga agtttattaa agtgttaatt cacagcttta ttgcctgttt
     3421 tgagaatctg cttgcataag tgatttttaa acttgatctg attctcgttt gtttcgaagt
     3481 tcttcagctt tgaaagtcag atcttcattc acagccatgc catagtttta ggatacagta
     3541 taagtagacg aattgggatg catcgtagat tttatgctgt gtttgttatg gtttttcagt
     3601 ggtagagatt ttcagagtat tattagggtt atagtcagtg tcaggtttat ttatcgtcag
     3661 cgtgatgggc aaaaaacagc tggcagtacg ggctatgtct tcgtctagaa gaagacaaag
     3721 ccaactgctg ttactctcgc ccagaaatct gcgccaacat ggacacaccc agtggagcaa
     3781 gccatctgat tttcatggtc agattgtaaa gaagcaatta ctcttggaca gtttaagcag
     3841 aaaagccacc aagtcaagtc actggtcata tgatgaactc gaagatctaa taaaagttaa
     3901 tttaagcgtt cattacaata aattcaagtg aatctgtttt aagctagcct ttaacgtttt
     3961 taaattcctt caaacaaatt ggatcctctt tgcagttcct catttctcgg tgagctggtg
     4021 ctttaatccc tttacttttt cttcctctag ggagtgatgg ttggcatggg acgaattaat
     4081 tcctgcagcc cgggggtacc ggtcgccacc atgaacaccc cgggaattaa cctgatcaag
     4141 gaggacatgc gcgtgaaggt gcacatggag ggcaacgtga acggccacgc cttcgtgatc
     4201 gagggcgagg gcaagggcaa gccctacgag ggcacccaga ccgccaacct gaccgtgaag
     4261 gagggcgccc ccctgccctt cagctacgac atcctgacca ccgccgtgca ctacggcaac
     4321 cgggtgttca ccaagtaccc cgaggacatc cccgactact tcaagcagag cttccccgag
     4381 ggctacagct gggagcgcac catgaccttc gaggacaagg gcatctgcac catccgcagc
     4441 gacatcagcc tggagggcga ctgcttcttc cagaacgtgc gcttcaaggg caccaacttc
     4501 ccccccaacg gccccgtgat gcagaagaaa accctgaagt gggagcccag caccgagaag
     4561 ctgcacgtgc gcgacggcct gctggtgggc aacatcaaca tggccctgct gctggagggc
     4621 ggcggccact acctgtgcga cttcaagacc acctacaagg ccaagaaggt ggtgcagctg
     4681 cccgacgccc acttcgtgga ccaccgcatc gagatcctgg gcaacgacag cgactacaac
     4741 aaggtgaagc tgtacgagca cgccgtggcc cgctacagcc ccctgcccag ccaggtgtgg
     4801 taatctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4861 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     5041 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5101 ttttttaacc aataggccga aatcggcgga tcctgcggcc gcaagggaaa atagaatgaa
     5161 gtgatctcca aaaaataagt actttttgac tgtaaataaa attgtaagga gtaaaaagta
     5221 cttttttttc taaaaaaatg taattaagta aaagtaaaag tattgatttt taattgtact
     5281 caagtaaagt aaaaatcccc aaaaataata cttaagtaca gtaatcaagt aaaattactc
     5341 aagtacttta cacctctggt tcttgacccc ctaccttcag caagcccagc agatccacta
     5401 gatcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     5461 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5521 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5641 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5701 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5761 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5941 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6361 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6421 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6601 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6661 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6721 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6781 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6841 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6901 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6961 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7021 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7081 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7141 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7201 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7261 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7321 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7381 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7441 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7501 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7561 acaaataggg gttccgcgca catttccccg aaaagtg
//
