LOCUS       sequence-212744-        4796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_799 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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_799
FEATURES             Location/Qualifiers
     source          1..4796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     129..146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     130..146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        173..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          818..914
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(975..977,978..980,981..1691)
                     /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(1020..1041)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1281..1300)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1628..1649
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2081..2215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2105..2124)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2159..2178
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2326..2342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2326..2342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2339..2361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2350..2366
                     /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(2374..2380,2381..2398,2399..2404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2419..2440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2557..2574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2728..3316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2808..2827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3487..4278,4279..4347))
                     /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     4110..4129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4348..4452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4520..4538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4576..4598)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4698..4717
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
       61 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      121 cacgacgttg taaaacgacg gccagtgagt tcgagcttgc atgcctgcag gtcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagaggatc
      781 cggtactaga ggaactgaaa aaccagaaag ttaactggta agtttagtct ttttgtcttt
      841 tatttcaggt cccggatccg gtggtggtgc aaatcaaaga actgctcctc agtggatgtt
      901 gcctttactt ctaggcctgt acggaagtgt tacttctgct ctaaaagctg cggaattgta
      961 cccgcgggcc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1021 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1081 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1141 tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc
     1201 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1261 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1321 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1381 acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg
     1441 acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca
     1501 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1561 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     1621 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1681 agctgtacaa gcgaattggt ggagggggcg gttccggtgg cggaggttcc cgaattcgtc
     1741 gacggagatc tgatatgttg cattttgcgt ggctgcttcc gcatttgcct gtgattcact
     1801 gcaataaaga cgattctcag gaagcgtcct cgcagactcc gtttttgcca ctcggctttg
     1861 ctttcaccac gttgtatatc tgcctgacta tctacagcgg aggtgatgat gaaagtatgg
     1921 atggcgatat taataagtat cttgtgctgg caatcatcgg cgtgggcggt ctgtccggcc
     1981 tggtcgccag ccagagcaca ggaagaaatt tcccgcctgc aacagcaaat aaaacagtgg
     2041 gacgatgaat cggtaccctc gagaagcttg cggccgcggg gatccagaca tgataagata
     2101 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2161 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2221 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttcgga
     2281 tcctctagag tcgatctgca ggcatgctag cttggcgtaa tcatggtcat agctgtttcc
     2341 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2401 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2461 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2521 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4501 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4561 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4621 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4681 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4741 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccat
//
