LOCUS       sequence-457269-        4229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for 2xHA GPR153.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4229)
  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 4229)
  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 GPR153
FEATURES             Location/Qualifiers
     source          1..4229
                     /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(2458..2557)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2669..2688)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2680..3489
                     /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(2747..2766)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3579..4167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4068..4087
                     /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 caaccatgtc
      601 agacgaaagg agacttcctg gaagtgcggt cgggtggctt gtatgcggtg gtctcagttt
      661 gctggccaat gcttggggta tactgtctgt aggagcgaag cagaagaaat ggaagcccct
      721 tgaatttctg ctttgcacat tggcagctac ccacatgctc aacgttgcgg tgccgattgc
      781 gacttactcc gtggtgcaac ttcggcgcca aaggccagac ttcgaatgga acgaaggcct
      841 gtgcaaggtt ttcgtaagta ctttctatac attgactctt gctacatgct ttagcgtaac
      901 atctctgagt tatcatcgga tgtggatggt ctgttggccg gtcaactacc gcctgtccaa
      961 cgctaaaaaa caggcggtac atacggtaat ggggatatgg atggtttcct tcatactctc
     1021 cgcactccca gccgttggat ggcacgatac gtcagaacga ttttacactc acgggtgtcg
     1081 atttatcgtc gccgaaatag gcctcggttt cggtgtatgt tttttgcttc tggttggggg
     1141 gagcgtcgcc atgggggtaa tctgcacagc gatcgcactt ttccaaacac ttgcagtcca
     1201 agtcggacgc caagccgata gacgagcctt cactgttccc acaatagtcg tggaagatgc
     1261 gcaagggaaa cgaagatcca gcatcgacgg cagcgaacct gcgaagacgt ctctccaaac
     1321 gacgggcctc gtgacaacaa ttgtttttat ctacgactgt ctcatggggt tcccggtttt
     1381 ggttgtgtca ttcagcagcc tcagggcgga tgcgtcagcg ccgtggatgg ccctttgtgt
     1441 tctttggtgc agcgttgcgc aagcgttgtt gctcccagta ttcctgtggg cttgcgatag
     1501 gtacagagca gatctgaaag ctgtgagaga gaagtgcatg gcgttgatgg caaatgacga
     1561 ggagtccgat gacgaaacca gtcttgaagg aggcataagc cctgatcttg tattggagcg
     1621 atcactcgat tacggatatg gaggtgactt cgtggcgctt gatcgcatgg caaagtacga
     1681 gatatctgca cttgaggggg gacttcctca actgtaccca ctccggccct tgcaggaaga
     1741 taaaatgcag tacctgcagg ttcctccaac gaggcggttc agtcacgacg atgcggatgt
     1801 atgggctgca gtcccccttc ctgcattctt gcccagatgg gggtccgggg aggacttggc
     1861 cgcactggct cacttggttc ttccagccgg acctgaacgg agacgagctt ctcttctcgc
     1921 ctttgctgag gatgcgcccc cgtcacgcgc acgacggaga tcagccgaaa gcctgctttc
     1981 cctgcgacct agtgctctcg attctgggcc cagaggagcc cgggactccc cgcctgggtc
     2041 tccgagacgc cggcctggtc ctgggccgcg gagcgcaagc gcgagccttc ttcctgatgc
     2101 tttcgccctt actgcatttg agtgtgagcc gcaagcattg aggcggccgc ccggtccatt
     2161 tcccgccgca cctgcggcac ctgacggggc agatccaggt gaggccccaa cccccccgtc
     2221 atctgcgcag agaagtcctg gtccgcggcc ctcagcgcat agccacgctg ggtcacttcg
     2281 acctggactg tcagccagtt ggggagagcc aggtggcctt cgagccgccg ggggaggagg
     2341 gagtacttct tcttttttga gtagtccgag tgagtcttcc ggctacgcca ctttgcatag
     2401 tgactctctt ggttccgcaa gttagtaaga attcgcggcc gcactcgaga tatctagacc
     2461 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2521 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gctctggccc
     2581 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2641 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2701 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2761 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2821 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2881 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2941 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3001 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gtccctgcgc
     3061 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     3121 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3181 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3241 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3301 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3361 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3421 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3481 tttttctaat cagaattggt taattggttg taacattatt cagattgggc cccgttccac
     3541 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3601 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3661 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3721 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3781 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3841 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3901 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3961 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4021 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4081 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4141 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4201 gccttttgct ggccttttgc tcacatgtt
//
