LOCUS       sequence-209362-        4865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_811 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4865)
  AUTHORS   Collins A, Huett A
  TITLE     A multi-phenotypic imaging screen to identify bacterial effectors by
            exogenous expression in a HeLa cell line.
  JOURNAL   Sci Data. 2018 May 15;5:180081. doi: 10.1038/sdata.2018.81.
  PUBMED    29762554
REFERENCE   2  (bases 1 to 4865)
  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"; journalName: "Sci Data.";
            date: "2018-05-15"; volume: "5:180081. doi"; pages: " 
            10.1038/sdata.2018.81"
COMMENT     Sequence Label: pCMV_mEm_4GS_LF82_811
FEATURES             Location/Qualifiers
     source          1..4865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     236..258
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     250..267
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     251..267
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        294..597
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..801
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     752..772
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     798..822
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          939..1035
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1096..1098,1099..1101,1102..1812)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1141..1162)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1402..1421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1749..1770
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2271..2405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2295..2314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2349..2368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2516..2532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2516..2532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2529..2551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2540..2556
                     /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)."
     promoter        complement(join(2564..2570,2571..2588,2589..2594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2609..2630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2747..2764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2918..3506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2998..3017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3677..4468,4469..4537))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4300..4319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4538..4642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4710..4728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4766..4788)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
       61 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
      121 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      181 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      241 tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca
      301 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      361 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      421 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      481 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      541 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      601 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      661 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      721 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      781 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
      841 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat
      901 ccggtactag aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt
      961 ttatttcagg tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt
     1021 tgcctttact tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt
     1081 acccgcgggc ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1141 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1201 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1261 gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt cgcccgctac
     1321 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1381 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1441 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1501 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta tatcaccgcc
     1561 gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat cgaggacggc
     1621 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1681 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     1741 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1801 gagctgtaca agcgaattgg tggagggggc ggttccggtg gcggaggttc ccgaattcgt
     1861 cgacggagat ctgatatggc tactacacca ttctattatg ccttttcagc gatatttgat
     1921 atcgtagcag tattaaatac cgaggaaata atcatgatat caggtcttaa tggtggaaat
     1981 ttcatcctta gctcgatggc taaggaaaga gtcaaatcag gtttacttga tagagcaaca
     2041 atagaaggtt ctaaaggcga gtataccgtg atttttgatg gttcagagtg ggaaggtaag
     2101 ccattagtct taagtagctc cagacagcct tatgaagcac gagtattcaa aagcatagat
     2161 ggtgcagctg cagaactact acgcataggc ttaagcgaag cttcaatcaa aaacaccacc
     2221 aaatcgaaca ataaatgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca
     2281 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2341 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2401 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2461 ttttttcgga tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat
     2521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4681 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     4741 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4801 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4861 gcggg
//
