LOCUS       sequence-209446-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_246 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  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 5285)
  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_246
FEATURES             Location/Qualifiers
     source          1..5285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        339..542
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     493..513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     539..563
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          680..776
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(837..839,840..842,843..1553)
                     /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(882..903)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1143..1162)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1490..1511
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2432..2566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2456..2475)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2510..2529
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2677..2693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2690..2712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2701..2717
                     /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(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2908..2925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3838..4629,4630..4698))
                     /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     4461..4480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4699..4803)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4871..4889
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4927..4949)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5049..5068
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5262..5284
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5276..5285,1..8)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(5277..5285,1..8)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
       61 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      181 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      241 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      301 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      361 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      421 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      481 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      541 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      601 gaagacaccg ggaccgatcc agcctccgga ctctagagga tccggtacta gaggaactga
      661 aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc
      721 cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct
      781 gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggg cccaccatgg
      841 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      901 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      961 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1021 tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1081 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1141 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1201 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1261 tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca
     1321 tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1381 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1441 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1501 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcgaattg
     1561 gtggaggggg cggttccggt ggcggaggtt cccgaattcg tcgacggaga tctgatatga
     1621 gaatactcgt tgctggcgca acaggaagta ttggtattca tgttgtgaat actgctattg
     1681 caatgggcca tcagcccgtg actctggtca ggaatcggcg gaaaatcaag ttactccctc
     1741 gtggaacaga tattttctat ggcgatgttt caatacctga aacacttacc gatttaccga
     1801 aagatattga tgccatcatt tttacactcg gctctgatgg tcagggacgt attggcgcca
     1861 gagcaataga ttacggcggt gtacgtaaca ttttgcgaat attcatggat acacctgttc
     1921 gcatcgccct gatgacaacg attggcgtaa ctgagcggct cagtagctgg aatcaacgta
     1981 ctgaagttca tgactggaag agacgctctg aacgcctggt cagagccagt gggcatatct
     2041 acactatcgt cagacccggc tggtttgatt acaacaatga tgatgagcac agaatcgtca
     2101 tgcttcaggg ggacagacgc catgcgggaa cgccggaaga tggtgttatc tcccgggagc
     2161 aaatcgccca ggttcttgta acagctctga gcaacgatgc ggcaaaaaat aaaacgttcg
     2221 aactggtcgc agagcgagga gaagcacagc aggatctcac tcctctgttt gcagagctgc
     2281 ggaatgataa tccacaaaaa aatgatgggg ttttcgatat agacaatatg ccccttactg
     2341 aagaacctga gtgcgttatt aacgacctga atctgtattc gaaaaattca aaaatctgaa
     2401 tcggtaccct cgagaagctt gcggccgcgg ggatccagac atgataagat acattgatga
     2461 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2521 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2581 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcgg atcctctaga
     2641 gtcgatctgc aggcatgcta gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4801 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4861 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4921 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4981 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5041 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5101 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5161 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5221 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5281 aacga
//
