LOCUS       sequence-471979-        3348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible lentiviral vector for 2xHA GPR153 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3348)
  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 3348)
  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: pInducer20 2xHA GPR153
FEATURES             Location/Qualifiers
     source          1..3348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(294..312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        380..484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(485..553,554..1345)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(703..722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2258..2275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2515..2536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2551..2556,2557..2574,2575..2581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2589..2605
                     /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)."
     primer_bind     2594..2616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2613..2629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2613..2629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2679..2698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(2890..3348,1..175)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
       61 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      121 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
      181 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
      241 gagaggcccg ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt
      301 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      361 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      421 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      481 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      541 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      601 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      661 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      721 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      781 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      841 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      901 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      961 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1021 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1081 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1141 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1201 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1261 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1321 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2221 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2281 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2341 caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2401 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2461 cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2521 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2581 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     2641 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2701 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
     2761 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
     2821 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
     2881 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
     2941 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
     3001 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
     3061 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
     3121 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
     3181 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
     3241 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
     3301 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtct
//
