LOCUS       sequence-209617-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_686 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  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 5912)
  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_686
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(829..831,832..834,835..1545)
                     /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(874..895)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1135..1154)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1482..1503
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3051..3185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3075..3094)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3129..3148
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3296..3312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3309..3331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3320..3336
                     /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(3344..3350,3351..3368,3369..3374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3389..3410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3527..3544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3698..4286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3778..3797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4457..5248,5249..5317))
                     /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     5080..5099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5318..5422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5490..5508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5546..5568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5668..5687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5881..5903
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5895..5912
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5896..5912
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag
      841 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      901 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      961 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1021 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1081 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1141 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1201 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1261 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
     1321 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1381 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1441 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1501 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgaat tggtggaggg
     1561 ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga gatctgatat gattggcgtg
     1621 aagtcctgcg gtggtcctaa gcagacgccg gattcggcgg ataataccac tgttgccgac
     1681 ctgactcagg aggaactgcg cattctcggc atcgagggcg acacgccgca ggatacgctg
     1741 cgtacgctgg ttggacgtat taaaaccatt cagacgaatc aggaaaatct ggagaaacag
     1801 aatgttgcac tggcagaaga aaatgcccgg ttgcaaaaca agggcaataa cgtggacggg
     1861 cgcatcaatg atgcggtgaa tgccgccaaa caggagagta cccaggcgca gcaaaaactt
     1921 caggctgaac agcagcggct ggctggcctg ctggaatcac tgaaaaatac cgttcccggc
     1981 ccgggagcgg gcagcaatac ttccgcaggc agtgacttac ctgtcgggct ggggctgggg
     2041 ctggaagatg ccgggcaggc cgaaacggtg gcgcagggta cgctctggat tgaaccgcag
     2101 gatggtgtac aactggatcg caacggcaaa ccggtgaccg gtgcagcagc tgcgaccaat
     2161 ggcactggca cattccagtt ccccacggcg ttcggtgggc tgaatgacaa cgcgatcact
     2221 cgtcagaaag ccgaactgga tcgtaatgcg aaaaatcagc aggccattga ggaaaccgat
     2281 ctgccggttt ataccctgcc ggaaaactcc acgctggttg gtagccaggc catgaccgcc
     2341 ctgctggggc gcgtgccgat tgatggcaaa gtcaccgacc cctatccctt taaagtgctt
     2401 attggtaaag agaacctgac cgccaacggc atcgagctgc cagatgtcga aggcgcgatt
     2461 gtgtccggta cggccaccgg cgactgggtg ttgtcctgtg tacgcggtga agttcactcc
     2521 attacgtttg tcttcactga tggcacggta cgcacggtac cgcagtcggc aaaatccagc
     2581 gatggcagca atggaaaatc caatggcggc agtattggct ggttgtccga tgagcacggt
     2641 attccgtgcc tgtccggcga ccgcaaaagc aacgcctcaa cttacctccc cacgttattt
     2701 gcgctgtctg ctgccggtgc ggcctctgat gtgatgtccg aaagccagac cacgaccacc
     2761 actgacggaa cgtccatcac cagtacgttg accggcagcg ccggacaggc tgcgctgggc
     2821 aaggcgttat ccggcggcac gaacgagctg gcggaatggg tgaaagcgcg atacggccag
     2881 atgtttgatg cgatttacgt tcctcccggc gcgaatgttg ctgtgcatat tacccgccag
     2941 atcccgattg attacgaaga gaagggccgc aaggtcaaat acgattttga tgtcagccag
     3001 caacacgagc tggattgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca
     3061 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3121 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3181 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     3241 ttttttcgga tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat
     3301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5461 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5521 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5581 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5641 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5701 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     5761 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5821 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5881 cccagtcacg acgttgtaaa acgacggcca gt
//
