LOCUS       sequence-422965-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of barcoded reporter plasmid for PRESTO-Salsa; 
            EGFP/BC293.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  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 5114)
  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-BC293-P4B1
FEATURES             Location/Qualifiers
     source          1..5114
                     /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    2626..2760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2650..2669)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2704..2723
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3123..3141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3209..3313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3314..3382,3383..4174)
                     /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(3532..3551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4345..4933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4834..4853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5087..5104
                     /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 gacaggtcaa gaattcgata tcaacttgtc gacctagttc tagaggatcc agacatgata
     2641 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2701 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2761 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2821 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctgc aagcctcgtc
     2881 gtcctggccg gaccacgcta tctgtgcaag gtccccggcc ccggacgcgc gctccatgag
     2941 cagagcgccc gccgccgagg cgaagactcg ggcggcgccc tgcccgtccc accaggtcaa
     3001 caggcggtaa ccggcctctt catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc
     3061 cccctggcgg acgggaagta tccagctcga ccaagctggc cgcgtcgagt gaagacgaaa
     3121 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3181 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3481 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3541 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3601 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3721 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3841 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4141 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4201 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4261 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4321 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4381 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4441 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4801 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4861 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4921 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4981 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5041 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5101 cgaggaagcg gaag
//
