LOCUS       sequence-451545-        5967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression red fluorescent OXER1 protein in zebrafish larvae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5967)
  AUTHORS   Ma Y, Hui KL, Gelashvili Z, Niethammer P
  TITLE     Oxoeicosanoid signaling mediates early antimicrobial defense in 
            zebrafish.
  JOURNAL   Cell Rep. 2023 Jan 31;42(1):111974. doi: 
            10.1016/j.celrep.2022.111974. Epub 2023 Jan 10.
  PUBMED    36640321
REFERENCE   2  (bases 1 to 5967)
  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: "Cell Rep.";
            date: "2023-01-31"; volume: "42(1):111974. doi"; pages: " 
            10.1016/j.celrep.2022.111974. Epub 2023 Jan 10"
COMMENT     Sequence Label: pCS2-OXER1-mKate2
FEATURES             Location/Qualifiers
     source          1..5967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      92..101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1271..1981
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKAVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRAD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKL"
     CDS             1271..1981
                     /codon_start=1
                     /product="monomeric photoactivatable red fluorescent 
                     protein (Gunewardene et al., 2011)"
                     /label=PAmKate
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKAVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRAD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKL"
     polyA_signal    2019..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2043..2062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2097..2116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2266..2286)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2266..2284)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2305..2321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2318..2340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2329..2345
                     /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(2353..2359,2360..2377,2378..2383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2398..2419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2536..2553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2707..3295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2787..2806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3466..4257,4258..4326))
                     /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     4089..4108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4327..4431)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4457..4912)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4594..4615)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4806..4825
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4983..5967
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tgccaccatg gaactccaca acctttccag
      121 cccctctcca tccctttcca gttctgtgct cccgcccagt tttagtccat ctccgagtag
      181 cgccccatcc gctttcacca cagtgggagg atctagcggc ggcccatgcc accccacttc
      241 atcaagcctc gtgagcgcct ttcttgcccc tatccttgcc ctggagttcg tactgggcct
      301 cgtcggtaac tccctggccc tcttcatctt ctgcatccat acgagaccct ggacctcaaa
      361 cacagtcttc ctggttagcc tggtggcagc tgattttctg ctcatttcaa acctgccttt
      421 gcgagtggat tactatcttt tgcacgagac ctggaggttc ggtgcagcag cctgcaaagt
      481 caacctgttc atgctgtcca ccaatcggac agctagcgtg gtctttctga ccgcaattgc
      541 tttgaacaga tatcttaagg tcgtacagcc tcatcacgtc cttagccgag cgtccgttgg
      601 cgcagcagca agagtggccg gtgggctgtg ggtaggcatt cttctcctta atgggcacct
      661 ccttctttct accttcagtg ggccatcctg cctgagctat cgggttggca cgaaaccttc
      721 tgcgtccctg cgctggcacc aggccctgta cctgctggag ttttttctgc cattggcgct
      781 cattttgttc gccatcgtct ccattgggct cacaatccgc aatcggggtc tcgggggcca
      841 agccgggcca cagcgcgcca tgagggtgct ggcaatggtg gttgccgtct acactatttg
      901 ttttctgcct agcatcattt ttggaatggc aagcatggtg gcattttggc tgtctgcttg
      961 tagaagtctt gatttgtgta cccagctgtt ccatggttca ttggctttca cctacctgaa
     1021 ctcagttctg gatcccgttc tgtactgctt ctcaagtcct aacttccttc accagagccg
     1081 cgccctgctc ggcctgacaa gggggcggca gggcccagtg tccgatgaat cctcatacca
     1141 gcccagccgg cagtggaggt acagagaggc tagtcggaag gccgaggcca tcggcaaact
     1201 gaaggtgcag ggggaagtga gccttgaaaa agaaggaagt agccaaggcg gaggaattcg
     1261 tcctggatct atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct
     1321 gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa
     1381 gccctacgag ggcacccaga ccatgagaat caaggccgtc gagggcggcc ctctcccctt
     1441 cgccttcgac atcctggcta ccagcttcat gtacggcagc aaaaccttca tcaaccacac
     1501 ccagggcatc cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt
     1561 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
     1621 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat
     1681 gcagaagaaa acactcggct gggaggcctc caccgagacc ctgtaccccg ctgacggcgg
     1741 cctggaaggc agagccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa
     1801 cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta
     1861 ctatgtggac agaagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca
     1921 gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc acaaactgta
     1981 actcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     2041 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2101 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2161 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     2221 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     2281 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2341 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     2401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4081 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4441 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4501 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4561 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4621 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4681 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4741 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4801 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4861 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4921 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4981 cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     5041 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     5101 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc
     5161 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     5221 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     5281 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     5341 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     5401 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     5461 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     5521 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     5581 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     5641 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     5701 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     5761 gtacattact attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg
     5821 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     5881 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     5941 tatataagca atgctcgttt agggaac
//
