LOCUS       sequence-421303-        5115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of barcoded reporter plasmid for PRESTO-Salsa; EGFP/BC99.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  AUTHORS   Chen H, Rosen CE, Gonzalez-Hernandez JA, Song D, Potempa J, Ring AM,
            Palm NW
  TITLE     Highly multiplexed bioactivity screening reveals human and 
            microbiota metabolome-GPCRome interactions.
  JOURNAL   Cell. 2023 Jul 6;186(14):3095-3110.e19. doi: 
            10.1016/j.cell.2023.05.024. Epub 2023 Jun 14.
  PUBMED    37321219
REFERENCE   2  (bases 1 to 5115)
  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.cell.2023.05.024"; journalName: "Cell"; date: "2023-07-6-
            6"; volume: "186"; issue: "14"; pages: "3095-3110.e19"
COMMENT     Sequence Label: Salsa-BC99-P2A3
FEATURES             Location/Qualifiers
     source          1..5115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     370..389
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    388..443
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     489..508
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          644..1216
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        complement(1437..1475)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1508..1526
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1550..1568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1592..1610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1634..1652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1676..1694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1837..1853
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1837..1853
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(1861..1863,1864..1866,1867..2580)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1861..1863,1864..1866,1867..2577)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1906..1927)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2167..2186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2514..2535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2627..2761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2651..2670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2705..2724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3124..3142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3210..3314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3315..3383,3384..4175)
                     /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(3533..3552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4346..4934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4835..4854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5088..5105
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtcgag
       61 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
      121 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
      181 acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac
      241 agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc
      301 cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg
      361 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc
      421 aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc
      481 cccataattt ttggcagagg gaaaaagatc gatctcagtg gtatttgtga gccagggcat
      541 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc
      601 caaaatgatg agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga
      661 aggcatgaac atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct
      721 caaacagtgt atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt
      781 tgtttctacc aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac
      841 cttatcattt aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac
      901 cctgattgcc ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa
      961 aagtgaattt aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa
     1021 atgaaaagaa aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa
     1081 attatcatga gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga
     1141 aaactttgcc ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa
     1201 gggtccccaa actcaccctg aagttctcag ctctagagat atcgtcgagg ccgcgctagc
     1261 gtttaaactg tcgaccttaa ttaagagatc tgctcgaggc gcgtcagctg actagaggat
     1321 ccccgggtac cgagctcgaa ttcggggccg cggaggctgg atcggtcccg gtgtcttcta
     1381 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     1441 ctgcttatat aggcctccca ccgtacacgc ctacctcgac ccgggtaccg agctcgactt
     1501 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
     1561 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
     1621 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     1681 atcactgata gggagtggta aactcgacct atataagcag agctcgttta gtgaaccgtc
     1741 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     1801 ccagcctccg cggccccgaa ttcctgcagc ggccgctcta gaactagtgg atccccgcgg
     1861 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1921 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1981 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2041 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2101 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2161 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2221 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2281 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2341 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2401 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2461 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2521 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2581 agagagccgc gaattcgata tcaagcttgt cgacctagtt ctagaggatc cagacatgat
     2641 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2701 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2761 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2821 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctg caagcctcgt
     2881 cgtcctggcc ggaccacgct atctgtgcaa ggtccccggc cccggacgcg cgctccatga
     2941 gcagagcgcc cgccgccgag gcgaagactc gggcggcgcc ctgcccgtcc caccaggtca
     3001 acaggcggta accggcctct tcatcgggaa tgcgcgcgac cttcagcatc gccggcatgt
     3061 ccccctggcg gacgggaagt atccagctcg accaagctgg ccgcgtcgag tgaagacgaa
     3121 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3181 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3241 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3301 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3361 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3421 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3481 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3541 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3601 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3661 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3721 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3781 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3841 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3901 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3961 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4021 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4081 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4141 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4201 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4261 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4981 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5041 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5101 gcgaggaagc ggaag
//
