LOCUS       sequence-457267-        3563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for 2xHA HCAR3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3563)
  AUTHORS   Pedro MP, Lund K, Kang SWS, Chen T, Stuelten CH, Porat-Shliom N, 
            Iglesias-Bartolome R
  TITLE     GPCR Screening Reveals that the Metabolite Receptor HCAR3 Regulates 
            Epithelial Proliferation, Migration, and Cellular Respiration.
  JOURNAL   J Invest Dermatol. 2024 Jun;144(6):1311-1321.e7. doi: 
            10.1016/j.jid.2023.12.002. Epub 2023 Dec 14.
  PUBMED    38103827
REFERENCE   2  (bases 1 to 3563)
  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"; doi: 
            "10.1016/j.jid.2023.12.002"; journalName: "J Invest Dermatol"; date:
            "2024-06"; volume: "144"; issue: "6"; pages: "1311-1321.e7"
COMMENT     Sequence Label: pENTR1A 2xHA HCAR3
FEATURES             Location/Qualifiers
     source          1..3563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             512..538
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             542..568
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(1792..1891)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2003..2022)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2014..2823
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2081..2100)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2913..3501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3402..3421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt accgctagcg ccgccaccat gtacccatac gatgttcctg actatgcggg
      541 ctatccctat gacgtcccgg actatgcagg atccggagat ccacctgtcg caaccatgaa
      601 ccggcaccac ctccaggacc acttcctgga gattgacaaa aagaactgct gtgtgttcag
      661 ggacgatttc atcgccaagg tactcccccc tgtgcttggg ttggagttca tcttcgggct
      721 gcttggcaat gggcttgcat tgtggatttt ttgcttccat cttaagtcat ggaagtccag
      781 tcggattttc ttgttcaatt tggccgtagc ggatttcctc ctcattatct gccttccctt
      841 tgtaatggat tactacgtga gacgctctga ttggaaattc ggggatatcc cgtgccggct
      901 ggtactcttc atgtttgcga tgaacaggca agggagcatc attttcttga ctgtggtcgc
      961 cgttgatcgc tatttcagag tggttcatcc tcaccacgct cttaataaga tatccaactg
     1021 gacagcagct attatctcat gtttgttgtg ggggataacg gtcgggctga ctgtccatct
     1081 tttgaagaaa aagttgttga tacagaatgg gacggctaat gtatgtatat ccttctctat
     1141 ttgccatacg tttcgctggc acgaagctat gtttctcttg gaatttttcc tgcctctggg
     1201 tataatcttg ttctgctccg ccagaataat ttggtcattg agacagagac agatggacag
     1261 gcacgcgaag attaaacggg caatcacgtt cataatggtg gtagcaatcg tatttgtcat
     1321 ctgcttcctg ccgtcagtgg ttgtcaggat tcacatcttc tggctcctgc acaccagcgg
     1381 gactcagaac tgcgaagtat atcgatcagt ggacctggca ttcttcatca cgctcagttt
     1441 tacgtacatg aacagcatgc ttgaccctgt cgtgtattat ttttcaagcc cctcattccc
     1501 gaactttttt agcacactga ttaatcgctg cctccaaagg aagataacag gcgaacccga
     1561 taataaccgg tccacctctg tggaactcac gggagacccc aacaagacca ggggtgcccc
     1621 cgaggcgctc attgcgaact ctggggaacc ctggtcccct agctaccttg gaccaacgtc
     1681 aaataatcat agtaaaaagg ggcactgtca tcaggagcca gccagtcttg aaaagcagct
     1741 tggctgttgt atcgaataat aagaattcgc ggccgcactc gagatatcta gacccagctt
     1801 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1861 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagctctg gcccgtgtct
     1921 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1981 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2041 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2101 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2161 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2221 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2281 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2341 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgtccct gcgccggttg
     2401 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2461 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2521 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2581 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2641 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2701 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2761 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2821 taatcagaat tggttaattg gttgtaacat tattcagatt gggccccgtt ccactgagcg
     2881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3541 tgctggcctt ttgctcacat gtt
//
