LOCUS       sequence-422948-        5116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of barcoded reporter plasmid for PRESTO-Salsa; 
            EGFP/BC232.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5116)
  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 5116)
  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-BC232-P3D4
FEATURES             Location/Qualifiers
     source          1..5116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     371..390
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    389..444
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     490..509
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          645..1217
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        complement(1438..1476)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1509..1527
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1551..1569
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1593..1611
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1635..1653
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1677..1695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1838..1854
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1838..1854
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(1862..1864,1865..1867,1868..2581)
                     /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(1862..1864,1865..1867,1868..2578)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1907..1928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2168..2187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2515..2536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2628..2762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2652..2671)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2706..2725
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3125..3143)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3211..3315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3316..3384,3385..4176)
                     /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(3534..3553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4347..4935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4836..4855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5089..5106
                     /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 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga
      241 cagcaggggg ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag
      301 ccagcatatg ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat
      361 gatttgccct cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg
      421 caatgaaaat aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt
      481 ccccataatt tttggcagag ggaaaaagat cgatctcagt ggtatttgtg agccagggca
      541 ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt gaattctttg
      601 ccaaaatgat gagacagcac aacaaccagc acgttgccca ggagctgtag gaaagagaag
      661 aaggcatgaa catggttagc agaggggccc ggtttggact cagagtattt tatcctcatc
      721 tcaaacagtg tatatcattg taaccataaa gagaaaggca ggatgatgac cagggtgtag
      781 ttgtttctac caataagaat atttccacgc cagccagaat ttatatgcag aaatattcta
      841 ccttatcatt taattataac aattgttctc taaaactgtg ctgaagtaca atataatata
      901 ccctgattgc cttgaaaaaa aagtgattag agaaagtact tacaatctga caaataaaca
      961 aaagtgaatt taaaaattcg ttacaaatgc aagctaaagt ttaacgaaaa agttacagaa
     1021 aatgaaaaga aaataagagg agacaatggt tgtcaacaga gtagaaagtg aaagaaacaa
     1081 aattatcatg agggtccatg gtgatacaag ggacatcttc ccattctaaa caacaccctg
     1141 aaaactttgc cccctccata taacatgaat tttacaatag cgaaaaagaa agaacaatca
     1201 agggtcccca aactcaccct gaagttctca gctctagaga tatcgtcgag gccgcgctag
     1261 cgtttaaact gtcgacctta attaagagat ctgctcgagg cgcgtcagct gactagagga
     1321 tccccgggta ccgagctcga attcggggcc gcggaggctg gatcggtccc ggtgtcttct
     1381 atggaggtca aaacagcgtg gatggcgtct ccaggcgatc tgacggttca ctaaacgagc
     1441 tctgcttata taggcctccc accgtacacg cctacctcga cccgggtacc gagctcgact
     1501 ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt tctctatcac
     1561 tgatagggag tggtaaactc gactttcact tttctctatc actgataggg agtggtaaac
     1621 tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc
     1681 tatcactgat agggagtggt aaactcgacc tatataagca gagctcgttt agtgaaccgt
     1741 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
     1801 tccagcctcc gcggccccga attcctgcag cggccgctct agaactagtg gatccccgcg
     1861 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1921 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1981 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2041 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2101 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2161 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2221 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2281 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2341 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2401 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2461 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2521 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2581 agacacgcgc cgaattcgat atcaagcttg tcgacctagt tctagaggat ccagacatga
     2641 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2701 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2761 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2821 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct gcaagcctcg
     2881 tcgtcctggc cggaccacgc tatctgtgca aggtccccgg ccccggacgc gcgctccatg
     2941 agcagagcgc ccgccgccga ggcgaagact cgggcggcgc cctgcccgtc ccaccaggtc
     3001 aacaggcggt aaccggcctc ttcatcggga atgcgcgcga ccttcagcat cgccggcatg
     3061 tccccctggc ggacgggaag tatccagctc gaccaagctg gccgcgtcga gtgaagacga
     3121 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3181 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3241 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3301 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3361 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3421 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3481 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3541 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     3601 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3661 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3721 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3781 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3841 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3901 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3961 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4021 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4081 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4141 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4201 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4261 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4321 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4381 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4441 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4501 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4561 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4621 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4681 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4741 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4801 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4861 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4921 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4981 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5041 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5101 agcgaggaag cggaag
//
