LOCUS       sequence-451548-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression red fluorescent hcar1-4 protein in zebrafish larvae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  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 5751)
  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-hcar1-4-mKate2
FEATURES             Location/Qualifiers
     source          1..5751
                     /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"
     CDS             1055..1765
                     /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             1055..1765
                     /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    1803..1937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1827..1846)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1881..1900
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2050..2070)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2050..2068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2089..2105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2089..2105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2102..2124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2113..2129
                     /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(2137..2143,2144..2161,2162..2167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2320..2337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2491..3079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2571..2590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3250..4041,4042..4110))
                     /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     3873..3892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4111..4215)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4241..4696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4378..4399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4590..4609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4767..5751
                     /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 agtaacatca cctcctgcat
      121 ggaacctcac tcctttattt cttccatttt aaagcccatt ttcatattgg agttaatttt
      181 ggggttgcct ggaaatgtgt ttgctctgtg gattttattt ttcaagtctc cttggaaagc
      241 ctgcaatgtg tatttcttca gcctggtaat agctgacctc ctactcctca ctgggatgcc
      301 tttccgcatc gattattttt ttcgtggtga agactggata tttggcgaat ccatttgccg
      361 ccttgtttta tatgtcatat cagtaaactc ctcagcaagc gtggctttca tgacaatttt
      421 agctgtggac cgcttcttca ggatccttta cccacaccac cgtatttgtc gaatgaatgt
      481 gaaacaagga ttaatgttgt ctagcatgac ttgggtgatt gttcttctcc ttcgtcttcc
      541 atctgccctc agcccggtcc ttacatcaca caaaaactct tcaaagctca catgccacag
      601 tttcctctca tggacctcac catcagttca gatgagaata tataattcag tacagctaat
      661 agaagtcctg gtggctttca ccttggtggt tttttgcttg gtgcgtgttt tctctaacac
      721 caacattcgg cagtctaagg gaccgcaccg cagggtgaag agggctgtga gattgctttt
      781 catttttgtg atcatctttg ttctctgttt tctacctacg gtcatctttg gcaacttcct
      841 tcaagtgttt ccttgctctg aatccctcat tgtgtgtctc catgcctctt tagctttatc
      901 ctacatgaac agtgcactgg atccagtcat ctactgccac attaacgctt ggtttcgtga
      961 caccctgaaa ggcaaatcca actcattggg aatgacgaag tttaagatga gtgcgaagac
     1021 aaacaaaaaa cctggaggaa ttcgtcctgg atctatggtg tctaagggcg aagagctgat
     1081 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     1141 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc
     1201 cgtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     1261 cagcaaaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     1321 tgagggcttt acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     1381 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1441 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     1501 gaccctgtac cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt
     1561 gggcgggggc cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa
     1621 gaacctcaag atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc
     1681 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     1741 tagcaaactg gggcacaaac tgtaactcga gcctctagaa ctatagtgag tcgtattacg
     1801 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1861 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1921 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1981 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     2041 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     2101 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     2161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     4261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4681 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4741 gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact
     4801 catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact
     4861 cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac
     4921 catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca
     4981 cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca
     5041 tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc
     5101 aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg
     5161 ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg
     5221 gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt
     5281 caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat
     5341 atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt
     5401 gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc
     5461 cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa
     5521 tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg
     5581 cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc
     5641 cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc
     5701 ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
