LOCUS       sequence-212845-        6740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitochondria localized Dendra2 in zebrafish neutrophils.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   Zhou W, Cao L, Jeffries J, Zhu X, Staiger CJ, Deng Q
  TITLE     Neutrophil-specific knockout demonstrates a role for mitochondria in
            regulating neutrophil motility in zebrafish.
  JOURNAL   Dis Model Mech. 2018 Mar 28;11(3). pii: dmm.033027. doi: 
            10.1242/dmm.033027.
  PUBMED    29590639
REFERENCE   2  (bases 1 to 6740)
  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."; date: "2018-03-28"; volume: "11(3). pii: dmm.033027. doi"; 
            pages: " 10.1242/dmm.033027"
COMMENT     Sequence Label: Tol2-lyzC-mito-Dendra2
FEATURES             Location/Qualifiers
     source          1..6740
                     /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)"
     CDS             3146..3220
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3254..3946
                     /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    4057..4178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4094..4113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4148..4167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4302..4489,4490..4501)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4556..4576)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4556..4574)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4595..4611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4595..4611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4608..4630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4619..4635
                     /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(4643..4649,4650..4667,4668..4673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4688..4709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4826..4843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4997..5585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5077..5096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5756..6547,6548..6616))
                     /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     6379..6398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6617..6721)
                     /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 tccaacaatg 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 gagatacatt tgattagaag agtttgactt gggttgatta
      781 tagcaaatag tggaacacaa acagattttg aaatattctt gcaaatttat ttattaaatc
      841 cattcaaata ttgtcatttt ttgtactgtg taatttatat tgatttgctc atacaatatt
      901 tacatttcgc ttaaacaata cactattatt tatatttaaa aatagctata tataaatcaa
      961 atgtatagct cttttttaca caattaatca aagctaaaat atttgtcaca caaatacagt
     1021 aagccacttc acaatgatat cttcaaaatc tgtgcttgac tgaatcattt catttattgt
     1081 tttgtaattc acattatacc ttacgttctc caggcacagt ttgaattgtt gataaatgaa
     1141 gataatacat ctgatttaat tagtctcctt tgtttataat aactttaatc taatttgata
     1201 gtaataattc cagcaataca tccattacaa taaaaatcgc ttttttattt tgcacattag
     1261 tttgccggta tttgtttaaa gtttcaccat acttttaaat catgtgctat ttgaaaaggg
     1321 ctgttatgac tggaattaaa tgtttttttt tatatcagtg ttttagattg aagtacaata
     1381 tttataggta aaattcacca gcatatgctt aaagatagcc atgctccagg tccttgtgtc
     1441 cttctaaggt ctgtttgtgg tttaagctgt ttgttgacac acgcttgtct tctccatttt
     1501 tctgtgaagt tattgtcagt ttattgcaca ctgggttgca ttaagctgca gttttttcac
     1561 aacagttttc cacattgtaa cagtcagtct ccaaattatt ccatcagtcc tcaaatttag
     1621 tcatttgagg ccatgaaaac acttaaaact acatgaaggt agtttggcat taggttttag
     1681 acttgttctt gtttggcata ttgcaacctg tgtaaaaatg atttgatttt tatttttttt
     1741 tttttttttt cagaaagaaa cggtccaaaa agagttatgt gaacatttat ccatgtgttt
     1801 aatgtttttt aaattaaaaa aaaacatttt tattaaacca cttgtgacta aacctttttt
     1861 agttattcta ttttgtgacg agtctgaatg gccaaagtac tcttgaatcc aaaatcttta
     1921 tagtaaaatg tttaatctta cttgcagtgt tgggtataat ttgtaacaaa gttgaattcc
     1981 atatctgctg agtgaaaaag taaagtaaga gattgctgta aatatttagg ttatttattt
     2041 attaatacaa tatagaaagt aacaaaacac aatgtaaaaa atcaataaag aaagtaaata
     2101 ttaaaagtaa tgataaacta atgatgtaaa caatattagc aacaaagttt gtacaattca
     2161 gggagggtaa agaatttcac ctaatagtca aactctgtat taaataatat aaaatctttg
     2221 ttcttagtct gaagaggaga ctgtgagaga aagcttactg gttgttaaat tagaaaattt
     2281 gtaacttgca gattaaaatg atttaaataa ttttataaag gccatgtttg aaacaaatgt
     2341 aaaaacataa taaatgttaa gaatttcaga tattgagatt ttacaattta aaaaaagcaa
     2401 taaagtaata gcagagacct acactcttaa agtaaattac ccaactctgc ttactagtaa
     2461 cttttggaat agttactagt atccattgaa caaatactag aatctaatta aattgtgtga
     2521 gtgtccaatg aaaagattct tgtatttaaa acagtaccag tatctatata tgtacaaaat
     2581 ttagtgctag caaaatgcaa ataaatacta ctgcatataa tggattgcat tgtaaatcga
     2641 ctcgccataa tacatttatt tcctattttt acttttaaca aaaaataagt tcactgagta
     2701 tttcatgtca ttaaatcgca ttaaacctct tgcatgcttt atgtacacta gtgcagttcg
     2761 ctaaatgacc accggtgtca ctaataacac gatcgtttct gaattctgca ttaaacgcca
     2821 tttatacgtt gctcttatta gtaaacaatt tctccatcat gagctcgacc agtattgcat
     2881 atttcatcct cttaaatcca gagttgtttc ttttaatcac tttctcttca tgataaaagc
     2941 agtttttggt tttcagaagt ttacaacaaa cccagtcagt tattgcctca gcaaacactg
     3001 gagcaactgc acacttgtga aaagcttctt gcgaaacaca aacctttctc tggacttata
     3061 aataaaggca ttcagcagct ccaccataca cctccactgg tcttgataaa gcagatatca
     3121 gcagtgatac aggtacctcg ccaccatgtc cgtcctgacg ccgctgctgc tgcggggctt
     3181 gacaggctcg gcccggcggc tcccagtgcc gcgcgccaag atccattcgt tgggggatcc
     3241 accggtcgcc accatgaaca ccccgggaat taacctgatc aaggaggaca tgcgcgtgaa
     3301 ggtgcacatg gagggcaacg tgaacggcca cgccttcgtg atcgagggcg agggcaaggg
     3361 caagccctac gagggcaccc agaccgccaa cctgaccgtg aaggagggcg cccccctgcc
     3421 cttcagctac gacatcctga ccaccgccgt gcactacggc aaccgggtgt tcaccaagta
     3481 ccccgaggac atccccgact acttcaagca gagcttcccc gagggctaca gctgggagcg
     3541 caccatgacc ttcgaggaca agggcatctg caccatccgc agcgacatca gcctggaggg
     3601 cgactgcttc ttccagaacg tgcgcttcaa gggcaccaac ttccccccca acggccccgt
     3661 gatgcagaag aagaccctga agtgggagcc cagcaccgag aagctgcacg tgcgcgacgg
     3721 cctgctggtg ggcaacatca acatggccct gctgctggag ggcggcggcc actacctgtg
     3781 cgacttcaag accacctaca aggccaagaa ggtggtgcag ctgcccgacg cccacttcgt
     3841 ggaccaccgc atcgagatcc tgggcaacga cagcgactac aacaaggtga agctgtacga
     3901 gcacgccgtg gcccgctaca gccccctgcc cagccaggtg tggtaatcta gatcataatc
     3961 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4021 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4141 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     4201 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4261 cgaaatcggc ggatcctgcg gccgcaaggg aaaatagaat gaagtgatct ccaaaaaata
     4321 agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa
     4381 atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc
     4441 cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct
     4501 ggttcttgac cccctacctt cagcaagccc agcagatcca ctagatcttt tgttcccttt
     4561 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4621 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4681 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4741 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4801 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4861 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4921 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5101 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5161 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5221 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5281 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5341 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5401 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5461 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5521 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5821 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5881 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5941 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6001 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6061 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6121 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6181 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6241 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6301 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6361 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6421 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6481 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6541 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6601 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6661 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6721 gcacatttcc ccgaaaagtg
//
