LOCUS       sequence-210481-        5438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_699 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5438)
  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 5438)
  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_699
FEATURES             Location/Qualifiers
     source          1..5438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          136..232
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(293..295,296..298,299..1009)
                     /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(338..359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(599..618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     946..967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2041..2175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2065..2084)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2119..2138
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2286..2302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2286..2302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2299..2321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2310..2326
                     /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(2334..2340,2341..2358,2359..2364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2379..2400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2517..2534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2688..3276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2768..2787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3447..4238,4239..4307))
                     /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     4070..4089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4308..4412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4480..4498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4536..4558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4658..4677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4871..4893
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4885..4902
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4886..4902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4929..5232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5233..5436
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5387..5407
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5433..5438,1..19)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
       61 acaccgggac cgatccagcc tccggactct agaggatccg gtactagagg aactgaaaaa
      121 ccagaaagtt aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt
      181 ggtggtgcaa atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac
      241 ggaagtgtta cttctgctct aaaagctgcg gaattgtacc cgcgggccca ccatggtgag
      301 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      361 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      421 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      481 caccttgacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      541 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      601 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      661 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      721 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
      781 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      841 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      901 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      961 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc gaattggtgg
     1021 agggggcggt tccggtggcg gaggttcccg aattcgtcga cggagatctg atatgacgaa
     1081 gaaaactacc aaccgcaaat cagccccacg gcaggatttg ttccagaccg ttaccgataa
     1141 gattattacg gcgctggaga acgggattac accctggcgt aaagcatggc aaaccggaaa
     1201 agagggttta cctgctaatg ccataactgg ccggaactac agcggtatta acgtgatgct
     1261 gttatggatc gatgctgccg ataagggatt tcattccaat cgctggctga ccttcaaaca
     1321 agcgctggat gccggtggta atgtccggaa aggtgaaaaa tcgacgcttg taacgctgtt
     1381 taaaccgttc cagaaggagg caacggataa cagcggccag ccggtgtttg atgataacgg
     1441 cgaggtggtc atgcgggatc gctgttttat gaccagcttc cacctcttca acgttgagca
     1501 gtgtgaaaac ctgcccgaca acctttaccc ggaaataccg gttttacccg acgtggaacg
     1561 tattgctcgg gcagaacaga tcgcaacttg tagcggcgta cctgtgatac atcgttatca
     1621 ggaccgtgca tattaccgcc cgggagttga ccacattatg atgccggaag ccgctcagtt
     1681 tgattcggcg gaagactact acagtacgct gttgcatgaa ctggtgcatt ccaccggcca
     1741 tgcttcgcga ctggcgcgtg aaggcattgt ttccacttcg aagaagttcg gcgatcctgt
     1801 ttatgggttt gaagagctga tagcggagat gggagcggcc tttttgtgtg cggaacttgg
     1861 catttatggc gacttgcaac atgagagcta tatcgcgtcg tggctgcaaa cgctgaagga
     1921 agacaatcgt gctgtatttc gtgcagctcg ctttgcccgg gaagcgtttg aatacctggt
     1981 gttgcgtgag caagcgctgg cagcatgaat cggtaccctc gagaagcttg cggccgcggg
     2041 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2101 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2161 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2221 gtgtgggagg ttttttcgga tcctctagag tcgatctgca ggcatgctag cttggcgtaa
     2281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4441 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4501 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4561 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4621 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4681 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4741 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4801 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     4861 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagttcga gcttgcatgc
     4921 ctgcaggtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4981 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5041 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5101 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5161 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     5221 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     5281 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     5341 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     5401 aggcgtgtac ggtgggaggt ctatataagc agagctcg
//
