LOCUS       sequence-210217-        5066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_667 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5066)
  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 5066)
  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_667
FEATURES             Location/Qualifiers
     source          1..5066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     79..96
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        123..426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        427..630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     581..601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     627..651
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          768..864
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(925..927,928..930,931..1641)
                     /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(970..991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1231..1250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1578..1599
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2301..2435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2325..2344)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2379..2398
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2546..2562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2559..2581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2570..2586
                     /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(2594..2600,2601..2618,2619..2624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2639..2660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2777..2794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2948..3536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3028..3047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3707..4498,4499..4567))
                     /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     4330..4349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4568..4672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4740..4758
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4796..4818)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4918..4937
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc atgcctgcag
      121 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      661 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      721 ctagaggatc cggtactaga ggaactgaaa aaccagaaag ttaactggta agtttagtct
      781 ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga actgctcctc
      841 agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct ctaaaagctg
      901 cggaattgta cccgcgggcc caccatggtg agcaagggcg aggagctgtt caccggggtg
      961 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1021 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1081 aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc
     1141 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1201 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1261 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1321 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat
     1381 atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc
     1441 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1501 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     1561 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1621 ggcatggacg agctgtacaa gcgaattggt ggagggggcg gttccggtgg cggaggttcc
     1681 cgaattcgtc gacggagatc tgatatgatc cgcaataaaa cagcttttac cggattgttc
     1741 gtcctgagta cctgcctgag tgggtgtgcg agtgttgaca gtgtcaacag tgcgataaac
     1801 cagattagcg gtgggatctt ttcagtcggt tccgcaccgg cagacagtgg tccggtggtg
     1861 ttgtcgaaag gcgtaccgca ggccagccac aaaatagcac aggaataccg tgttcccgtt
     1921 gatgttgata cggcagctgc gcgggtaaaa cgctactacc gctttatctc ggctgaagaa
     1981 gtagcacagc tacataacaa cggtacgcaa agtggagggt ggactgccgc ttcagtagcc
     2041 gatggggcat gggtatggga tgggcaaccc ggtagctact acaaaatggg gcgcgactgg
     2101 ggaagcgaaa atactggtga cagcattcag gtcgagctgg agaaaaacgg ttccggtagc
     2161 cgcatgtata tcacctttcg ttccgggctg gcctctcatg ccacggcggc ttataccagc
     2221 aaactgtttg agaatatgaa gagcgtggcg gaagggacga ttcgatgaat cggtaccctc
     2281 gagaagcttg cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa
     2341 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2401 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2461 tgtttcaggt tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgatctgca
     2521 ggcatgctag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2581 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2701 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4681 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4741 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4801 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4861 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4921 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4981 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     5041 ggcgatcggt gcgggcctct tcgcta
//
