LOCUS       sequence-210475-        5771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_705 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5771)
  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 5771)
  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_705
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          125..221
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(282..284,285..287,288..998)
                     /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(327..348)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(588..607)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     935..956
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2363..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2608..2624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2608..2624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2621..2643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2632..2648
                     /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(2656..2662,2663..2680,2681..2686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2701..2722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2839..2856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3769..4560,4561..4629))
                     /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     4392..4411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4630..4734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4802..4820
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4858..4880)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4980..4999
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5193..5215
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5207..5224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5208..5224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5251..5554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5555..5758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5709..5729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5755..5771,1..8)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
       61 gatccagcct ccggactcta gaggatccgg tactagagga actgaaaaac cagaaagtta
      121 actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa
      181 tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg gaagtgttac
      241 ttctgctcta aaagctgcgg aattgtaccc gcgggcccac catggtgagc aagggcgagg
      301 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      361 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      421 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttgacct
      481 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      541 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      601 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      661 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      721 acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca
      781 agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      841 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
      901 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      961 ccgccgggat cactctcggc atggacgagc tgtacaagcg aattggtgga gggggcggtt
     1021 ccggtggcgg aggttcccga attcgtcgac ggagatctga tatggcatta atcggagtgt
     1081 atgcggattg ggaaggcctg gatggtccgg agcggattgg ttatttacac agtcgccgca
     1141 ctcggacgcg tgagatattt gagtttgagt acgataaaaa ggcgcttgca gatccttcac
     1201 ttaatttcat tcagttagat cctgaaataa tgttgtatga gggagcacag tacccaattc
     1261 ctccaaagga taaatttggt gcatttagtg attcctgccc ggatcgttgg gggcgtatgc
     1321 taatgaagcg tcgttttgag agggatattc gagacgggtt atgtgataaa gactcccacc
     1381 tttatgagtc tgattatcta ctaggtgttc atgatcttta tcgggttggg gctcttcgat
     1441 acaaacgtga agacgctgga gagtttcttg ataatcgcat tgatgttgca gccccaccat
     1501 ttaccgaaat agcgagtctg gaaagggcta gccgggcaat tgaagaagat cctgataata
     1561 aagagctaat ggggcaaaaa tggttaagaa tgctaatcgc tccgggggga tcattagggg
     1621 ggacacgtcc gaaagccagt gttgttgatg aagctgggca tctctacatc gctaaatttc
     1681 ccagtgtaaa agatgagtat gatgttggtg gatgggaaat ggtcgttaat gcacttgctg
     1741 ttggctgtgg actgaatgtt gcacctgctc aggcgcataa atttgccagc aattaccatt
     1801 gttttatggt tcgtcgtttt gaccgcacta atgcgggaag gcgtctgcat tttgcttcgg
     1861 ctatgacact aaccaggcat caggatggtg aagatgcctc tacaggtgtg agttatctgg
     1921 aattagcaga tgttcttatc agacatggag cgcagactaa tactgacctt aaggaactct
     1981 ggtccaggat tgtttttaat attcttgttt ctaataccga tgaccattta cgtaatcatg
     2041 gatatatctt gataccggga aaaggttggc ggctttctga ggcatatgat ttgaatcctg
     2101 ttgccaggtc tgacggtcta aaactcaata ttacagaaaa tgataatgca cttgatttgg
     2161 aattagctcg agaagttgcg gagtattttc gacttggttt gacagaagca gatgacatta
     2221 ttgctaattt tagagggatt gtcagtcagt ggcgaatcat tgccgagcgt cttcgattac
     2281 ctggtcgtga gcaagagtta atggcagagg cgttcagggg ggcaatttga atcggtaccc
     2341 tcgagaagct tgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca
     2401 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgatctg
     2581 caggcatgct agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4741 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4801 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4861 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4921 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4981 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5041 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5101 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5161 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5221 cagtgagttc gagcttgcat gcctgcaggt cgttacataa cttacggtaa atggcccgcc
     5281 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5341 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5401 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5461 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5521 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     5581 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     5641 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     5701 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     5761 tttagtgaac c
//
