LOCUS       sequence-209535-        5408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_353 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5408)
  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 5408)
  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_353
FEATURES             Location/Qualifiers
     source          1..5408
                     /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    2547..2681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2625..2644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2792..2808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2792..2808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2805..2827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2816..2832
                     /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(2840..2846,2847..2864,2865..2870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2885..2906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3023..3040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3194..3782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3274..3293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3953..4744,4745..4813))
                     /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     4576..4595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4814..4918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4986..5004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5042..5064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5164..5183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5377..5399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5391..5408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5392..5408
                     /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 gaatattaaa
     1621 gtatcaatag ttatccctac ctttggtaga cctgagaatc ttcttagggc aatcagcagt
     1681 gtaatgagac aaacatataa taatttagaa attatcgttg ttgatgataa tggtaagggt
     1741 agcgaaaacc agattaaaac ggaagaattg cttaatgacg aaaagtataa agatataata
     1801 tacatagttt tacctaataa taaaggtggt ggaattgcta ggaattacgg tatagaaaaa
     1861 gcaagcggtg aagttattac atttttggat gatgatgatt attacttatg taataaagtt
     1921 gaagtccagt tacgtcatct aatagttaat aacttagata tatctttgtg cgatatgatt
     1981 attgaatctg atggttcatc acgattcaaa cattcaaaac gttattcaaa tgctaaagga
     2041 ataaatataa atgagttttt gattgatggt gtagctttta cacctatgat tatggtgaga
     2101 aaaaatatac tattaagtgc tggaggattt ttagatacac cacgttttca agatcatacg
     2161 ctgatgctaa agttattttt gattacggat aaggtcggtc atgttgaaag taaattattt
     2221 gtacattgtt ccaatatgaa tacaagagtt agtaactctc ccaaatcgtt gaaggggttt
     2281 ctaatacgac acagattaga aaactatgtg atgaagaacc aaaggaatat agtagagcaa
     2341 gttaaattca atcaatgcgc acaagtatct cctttccttt ataaaagaag gggttacaat
     2401 aaatatgttt ccttcatgat ctggtcttta aattataatc ttagtgtaaa aaatgtatta
     2461 ttaacatttg cgagggtttt aaaaatatca tgcttagcaa tattcaaaga ttgaatcggt
     2521 accctcgaga agcttgcggc cgcggggatc cagacatgat aagatacatt gatgagtttg
     2581 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2641 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2701 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct ctagagtcga
     2761 tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2821 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2881 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2941 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3001 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3061 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5161 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5221 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     5281 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5341 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5401 cggccagt
//
