LOCUS       sequence-210127-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_261 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  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 6875)
  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_261
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     186..208
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     200..217
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     201..217
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        244..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..772
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          889..985
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1046..1048,1049..1051,1052..1762)
                     /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(1091..1112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1352..1371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1699..1720
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4231..4365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4255..4274)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4309..4328
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4476..4492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4476..4492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4489..4511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4500..4516
                     /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(4524..4530,4531..4548,4549..4554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4569..4590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4707..4724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4878..5466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4958..4977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5637..6428,6429..6497))
                     /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     6260..6279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6498..6602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6670..6688
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6726..6748)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6848..6867
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
       61 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg catgcctgca
      241 ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
      781 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
      841 tctagaggat ccggtactag aggaactgaa aaaccagaaa gttaactggt aagtttagtc
      901 tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag aactgctcct
      961 cagtggatgt tgcctttact tctaggcctg tacggaagtg ttacttctgc tctaaaagct
     1021 gcggaattgt acccgcgggc ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     1081 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1141 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1201 caagctgccc gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt
     1261 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1321 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1381 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1441 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta
     1501 tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat
     1561 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1621 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     1681 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1741 cggcatggac gagctgtaca agcgaattgg tggagggggc ggttccggtg gcggaggttc
     1801 ccgaattcgt cgacgtagat ctgatatggc agtacagatt tcaggtgtgc tgaaagatgg
     1861 tgcgggaaaa ccaatacaga actgcactat tcaactgaag gcaaagcgta acagcaccac
     1921 ggtactggtg aacacggtgg cctctgaaaa tccggatgaa gccgggcgtt acagcatgga
     1981 tgttgagtat ggccagtaca gcgttatcct gcttgttgaa ggttttccgc cttcacatgc
     2041 cggaaccatt accgtctatg aaggttccag accaggtacg ctgaatgatt ttctcggtgc
     2101 catgacggaa gatgatgtca tgccggaggc attgcgtcgt tttgaggaaa tggtggaaga
     2161 agcggcacgc aacgctgaag ccgcctctca gagcgcagcg gcggcaaaga aatccgaaac
     2221 tgcagcggca tcatcgaaga acgcggcgaa aacctcagaa acgaatgcag ctaacagcgc
     2281 acaggcggca gcggcctcgc agactgcatc ggcaaactcc gcaacagcag ctaaaaaatc
     2341 agaaaccaac gcgaaaaata gcgagacagc cgcaaagacg agcgaaacca acgcaaagtc
     2401 cagccagacg gcagcgaaga ccagcgaaac gaatgctaaa gccagtgaaa ctgcggcaaa
     2461 aaacagccag gttgcagcag cccaaagcga gagcgcggca gccggttctg cgacttcagc
     2521 agctggatca gcaactgcta cggctaacag ccagaaagct gcgaagacga gtgaaactaa
     2581 cgcaaagtcc agccagacgg cagcgaagac cagcgaaacg aatgccaaag ccagtgaaac
     2641 tgcggcgaaa agcagtcagg atgcagcggc tcaaagcgag agtgccgcag ccagttctgc
     2701 aagcgcggcg gctgcttctg ccactgcatc agctaacagt caaaaagcag caaaaaccag
     2761 tgaaaccaac gcaaaggtga gcgaaacagc ggctgcgaac tcagcgaaag catcggcagc
     2821 aagccagacg gcagctaaag caagcgaaga tgcagccaga gagtacgcaa gccaggctgc
     2881 ggagccgtat aaatatgtct tacagccgct gcctgatgtg tggataccat ttaacgattc
     2941 gctggatatg attacgggct tttcgccgtc atataaaaaa attgttattg gtgacgatga
     3001 aataacgatg cctggcgaca agattgttaa gtttaaacgt gcatcgaaag caacctatat
     3061 taacaaatct ggtgtgctga cagaggctgc cattgacgag ccacgatttg aacgtgatgg
     3121 cctgcttatt gaggggcaaa gaacaaacta catgctcaat tcggaaagcc ctgccagttg
     3181 ggggcgatcg tcaaatatgg atgtgcccga aacagggacg gataattttg gttttaccta
     3241 tggaaagttt gtctgcaacg attctctgat tgggcaaacc tcagccatta atatggcatc
     3301 aattgctgca acaaagtcag ttgatgtctc aggcgataat aaacacgtga caacctcatg
     3361 tcgttttaaa acagaactgc aggtaaggtt gcgtatccgg tttgataaat atgacggtag
     3421 cgcaacaact tttcttggtg atgcgtatat tgatacacaa acgcttgaaa ttaatatgac
     3481 aggcggtgct gcctcaagga ttacagcgag agtcagaaag gacgaagcta ccggatggat
     3541 ttttgcagag gcaacaattc aggcaattga tggggagtta aaaataggct ctcagataca
     3601 gtattctcct aagcagagcg gggcaaccgt atctggtgac tatatttatc tggccacccc
     3661 acaagtagaa gatgggcctt gtgtatcatc ttttattata tcaggagcga cggcggcgac
     3721 ccgcgcaagc gatatagtta cagttccaat taagaataat ctttataatc ttccttttac
     3781 ggttctttgt gaggtacata agaactggta taaaacgcca aatgcagcgc cacgtgtttt
     3841 tgataccggc ggtcatcaaa ccggagcggc tattattctt ggcttcggtc gttcaacaga
     3901 ttacgacgga tttccttatt gcgatattgg aggagctaac agacgggtaa acgaaaacgc
     3961 atcgcttgaa aaaatggtta tggggatgcg tgtaaagtca gagcagtcta cgtgctcagt
     4021 aagtaacggg catatatcca gcgaaacaaa aaccacatgg tcctgtattc agaacaccgc
     4081 aattatccgt attggaggcc agactacagc cgggttacgt catttatttg gtcatgtcag
     4141 gaatttcaga atatggcaca aggcattgac tgatgctcag gtgggggagt caatctgaat
     4201 cggtaccctc gagaagcttg cggccgcggg gatccagaca tgataagata cattgatgag
     4261 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4321 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4381 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttcgga tcctctagag
     4441 tcgatctgca ggcatgctag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4501 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4561 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4621 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5761 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6601 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6661 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6721 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6781 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6841 aactatgcgg catcagagca gattgtactg agagt
//
