LOCUS       sequence-209418-        5306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_098 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5306)
  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 5306)
  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_098
FEATURES             Location/Qualifiers
     source          1..5306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     99..116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     100..116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        143..446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        447..650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     601..621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     647..671
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          788..884
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(945..947,948..950,951..1661)
                     /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(990..1011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1251..1270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1598..1619
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2561..2695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2585..2604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2639..2658
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2806..2822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2806..2822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2819..2841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2830..2846
                     /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(2854..2860,2861..2878,2879..2884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2899..2920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3037..3054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3208..3796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3288..3307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3967..4758,4759..4827))
                     /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     4590..4609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4828..4932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5000..5018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5056..5078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5178..5197
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
       61 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt
      121 tcgagcttgc atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac
      181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      421 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      481 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      541 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      601 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      661 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
      721 accgatccag cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag
      781 ttaactggta agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc
      841 aaatcaaaga actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt
      901 tacttctgct ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg
      961 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1021 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1081 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga
     1141 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1201 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1261 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1321 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1381 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact
     1441 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1501 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1561 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1621 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gcgaattggt ggagggggcg
     1681 gttccggtgg cggaggttcc cgaattcgtc gacggagatc tgatatgacc actatgacca
     1741 aagagcgact actgacaatc aagcagtggc gcgaaacata cggacctggt agcaacgttg
     1801 tactgccagc agaagaagcg gaagaactgg cacgaattgc tctggcatcg ctggaagcag
     1861 agcctattgg tgaggtttca gagaagcgac tcggccttgt tatggatgga acggtagacc
     1921 ttggcgggaa atcaacttat cgcatcatta agggagaaaa agcgatgaag ttgttgccgc
     1981 tggggacgaa gttttatacc gcccctccag tgccagtagt acctgaagaa aaaccaatgc
     2041 ctaaccctct taaaatgtac gcggtcgatg ctgttgccgc tattgcagag gtgagaggct
     2101 ggaactcctg ccgcgccgct atgcttcagg gtagccaacc tgtaagccaa acttacaagt
     2161 ttccagttaa tacaccttgc caggatgcgc cagcccatat ctggctgcaa acggctgtag
     2221 tatggccaga agatggcgag ttaagcgaat taacgtggtg cagccacaat cagcaccatg
     2281 atgacacgct atatgttcga gctgaccttg tgaatggcaa ctatccggtt actccggatg
     2341 gttggataag ctgtagtgag cgaatgccgg ataagttaat tccggtaatg gtcatgtatg
     2401 aagacggtga gatgtggtct gcaatgtgga atggcaatcg ctgggatgat ggcaccgaat
     2461 atccggatcc gcactcagtt acgcactggc gtgaaatgcc agcagcacca cagcaggagg
     2521 tgaagtgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca tgataagata
     2581 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2641 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2701 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttcgga
     2761 tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat agctgtttcc
     2821 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2881 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2941 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3001 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3061 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4981 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5041 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5101 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5161 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5221 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5281 gctgcgcaac tgttgggaag ggcgat
//
