LOCUS       sequence-209384-        5147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_321 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5147)
  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 5147)
  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_321
FEATURES             Location/Qualifiers
     source          1..5147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..86
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     78..95
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     79..95
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        122..425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     580..600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..650
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          767..863
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(924..926,927..929,930..1640)
                     /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(969..990)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1230..1249)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1577..1598
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2381..2515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2405..2424)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2459..2478
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2626..2642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2626..2642)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2639..2661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2650..2666
                     /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(2674..2680,2681..2698,2699..2704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2857..2874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3028..3616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3108..3127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3787..4578,4579..4647))
                     /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     4410..4429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4648..4752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4820..4838
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4876..4898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4998..5017
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
       61 tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg
      121 tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      541 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      601 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      661 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      721 tagaggatcc ggtactagag gaactgaaaa accagaaagt taactggtaa gtttagtctt
      781 tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa ctgctcctca
      841 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc
      901 ggaattgtac ccgcgggccc accatggtga gcaagggcga ggagctgttc accggggtgg
      961 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1021 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1081 agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg cagtgcttcg
     1141 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1201 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1261 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1321 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata
     1381 tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg
     1441 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1501 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1561 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1621 gcatggacga gctgtacaag cgaattggtg gagggggcgg ttccggtggc ggaggttccc
     1681 gaattcgtcg acggagatct gatatggcaa aaatattacc ttctatattt ggcgcggata
     1741 ttttacgcct gaaagatgaa attcattttc tggaagagga acaaactgaa atattgcacg
     1801 ttgatttaat ggatggcact tacgtcagta atattgcctt tggcccaaat caaattgcag
     1861 caatgaaaaa agccagttcg atgatctttg atgtgcatat gatgctggca aatcctgagc
     1921 gacatattga tgatgtaatc aataccgggg cggagatgat ttctgtacat tatgaatcta
     1981 cgccacatgt acattatatt attcaaaaaa tcaaaaaagc ggggcgcaaa gcaggtgttg
     2041 tgcttaatcc tgggacacct gaatctgtca tcgagtattt attagatgat attgattata
     2101 tattgttaat gacaattaac cccgggcaac caggccagac atttattcat aaaagtctgg
     2161 agaaaatccg taataccaaa aaaatgatat ctggcagaga tattcaaatt gaagtggatg
     2221 gtggcgttaa tcttgataat gtgaaacaag ttaaagaagc gggggctgac cttattgttg
     2281 ttggcggtgc attatttaac aaacagcctc atgaaagtta tcaggcatta aaaatcgctg
     2341 ttcagtgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca tgataagata
     2401 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2461 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2521 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttcgga
     2581 tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat agctgtttcc
     2641 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2701 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2761 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2821 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2881 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2941 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3001 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3061 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3121 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3181 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3241 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3301 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3361 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3421 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3481 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3541 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3601 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3661 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3721 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3781 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3841 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3901 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3961 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4021 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4081 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4141 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4201 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4261 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4321 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4381 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4441 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4501 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4561 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4621 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4681 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4741 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4801 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4861 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4921 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4981 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5041 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5101 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctat
//
