LOCUS       sequence-209728-        5192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_582 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  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 5192)
  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_582
FEATURES             Location/Qualifiers
     source          1..5192
                     /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    2331..2465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2355..2374)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2409..2428
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2576..2592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2576..2592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2589..2611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2600..2616
                     /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(2624..2630,2631..2648,2649..2654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2669..2690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2807..2824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2978..3566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3058..3077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3737..4528,4529..4597))
                     /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     4360..4379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4598..4702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4770..4788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4826..4848)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4948..4967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5161..5183
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5175..5192
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5176..5192
                     /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 ggcatttccc
     1621 gataactaca accgtctgtt tataccacgg caaaaaacta aagataataa tggcctcaaa
     1681 acactcaata gtaaagcggt tggcagactg cttgctgctg gcggcatata caatgggaat
     1741 atcgaaggat tcagggaaac agcaaacaag ttgggaggag atgcacctgc tggttacgat
     1801 caagttatgg ataacaaagg gttactcatc gctggtgctt caatagttgc tggagttgtt
     1861 ctgggaagaa tgagattccc tgatttggaa tcatttgagc atttcggagc cagaggagca
     1921 gtatcaggaa gaccatttaa tcccgacctt gccggtggtc caatagaaaa cctgacgata
     1981 gatggcgtaa caatcaatag ggaaggtatc gctattgttg aaaagcatat ctccagattt
     2041 gatcatgatc cagctaatga tgtaatgatt tcaaggctaa aaaaaatagc aaataaagaa
     2101 ttgcttcctg aaaaatacga tctgaactac tatactcatg aatgccgcga gtatcagcgt
     2161 tattgtaatt taggatggga aacaggtgaa ccgaatggtt tagatggata tgaattatgg
     2221 aataatgcac atactgcaac tctagaagac tttaaaatta aagatactga cttatttcat
     2281 ccggatgcaa aaaaatgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca
     2341 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2401 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2461 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2521 ttttttcgga tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat
     2581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2641 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4741 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     4801 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4861 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4921 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4981 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     5041 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5101 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5161 cccagtcacg acgttgtaaa acgacggcca gt
//
