LOCUS       sequence-209390-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_139 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  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 4922)
  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_139
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     182..204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     196..213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     197..213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        240..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        544..747
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     698..718
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     744..768
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          885..981
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1042..1044,1045..1047,1048..1758)
                     /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(1087..1108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1695..1716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2274..2408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2352..2371
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2519..2535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2519..2535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2532..2554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2543..2559
                     /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(2567..2573,2574..2591,2592..2597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2612..2633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2750..2767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2921..3509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3001..3020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3680..4471,4472..4540))
                     /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     4303..4322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4541..4645)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4713..4731
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4769..4791)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4891..4910
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
       61 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
      121 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
      181 tcccagtcac gacgttgtaa aacgacggcc agtgagttcg agcttgcatg cctgcaggtc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      721 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      781 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
      841 gaggatccgg tactagagga actgaaaaac cagaaagtta actggtaagt ttagtctttt
      901 tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt
      961 ggatgttgcc tttacttcta ggcctgtacg gaagtgttac ttctgctcta aaagctgcgg
     1021 aattgtaccc gcgggcccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accttgacct acggcgtgca gtgcttcgcc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa ggtctatatc
     1501 accgccgaca agcagaagaa cggcatcaag gtgaacttca agacccgcca caacatcgag
     1561 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1621 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     1681 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1741 atggacgagc tgtacaagcg aattggtgga gggggcggtt ccggtggcgg aggttcccga
     1801 attcgtcgac ggagatctga tatgaagagg attatcggcg tcgttgctgg cgctatattg
     1861 ttatctgggt gcgcaactat tgttggtgac gaaacacagc ttgtgcaagt gaacagcaat
     1921 ccttccggtg cgagctttaa ggtaaaagac gaatccggcg tgattgttgc gcaaggtaag
     1981 actccgcaag gagtaactct tgccaagtca gatggtagtt attttggcaa aaagagctac
     2041 cagatcacta tggaaaagga tgggtacgaa ccagttaccc tgccaatcaa agccaatgct
     2101 aatggttggt atattggtgg aaaccttgtg tttggtgggt taattggttg gcttgctgta
     2161 gatcctttta atggtgggat gtataccttg aagccaaaag aggcaaacgc atctcttata
     2221 ccatcaacaa agcaagactg aatcggtacc ctcgagaagc ttgcggccgc ggggatccag
     2281 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2341 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2401 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2461 aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt
     2521 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2581 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2641 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2701 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3361 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4681 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     4741 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4801 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4861 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4921 ac
//
