LOCUS       sequence-210850-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_677 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  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 4775)
  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_677
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     245..267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     259..276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     260..276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        303..606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        607..810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     761..781
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     807..831
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          948..1044
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1105..1107,1108..1110,1111..1821)
                     /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(1150..1171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1411..1430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1758..1779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2190..2324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2214..2233)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2268..2287
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2448..2470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2459..2475
                     /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(2483..2489,2490..2507,2508..2513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2528..2549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2666..2683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2837..3425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2917..2936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3596..4387,4388..4456))
                     /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     4219..4238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4457..4561)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4629..4647
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4685..4707)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
       61 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      121 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      181 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      241 ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc atgcctgcag
      301 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      361 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      421 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      481 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      541 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      601 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      661 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      721 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      781 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      841 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      901 ctagaggatc cggtactaga ggaactgaaa aaccagaaag ttaactggta agtttagtct
      961 ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga actgctcctc
     1021 agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct ctaaaagctg
     1081 cggaattgta cccgcgggcc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1141 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1201 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1261 aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc
     1321 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1381 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1441 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1501 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat
     1561 atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc
     1621 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1681 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     1741 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1801 ggcatggacg agctgtacaa gcgaattggt ggagggggcg gttccggtgg cggaggttcc
     1861 cgaattcgtc gacggagatc tgatatgatg aaacaacgtt atcgggttga agcggttatg
     1921 gcatcttcaa gacaaaataa gctggaagtc ccacgtgatg tgatggatgt tttgtgcgag
     1981 caggattgct cgtcactcca aatcccggaa attattgagc gtctgacttc gctgggctat
     2041 cgcccgagat atgaagccac agctgatacg ctcacagaca ttgtcaccct gtggatttgg
     2101 gtgggtcagg aggagatgtt gctgaactgc cagatggagc cgctggcggt gcattgaatc
     2161 ggtaccctcg agaagcttgc ggccgcgggg atccagacat gataagatac attgatgagt
     2221 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2281 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2341 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt
     2401 cgatctgcag gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2461 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2521 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2581 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2641 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2701 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2761 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2821 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2881 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2941 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3001 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3061 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3121 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3181 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3241 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3301 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3361 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3421 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3481 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3541 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3721 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3841 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3901 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4021 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4141 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4201 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4261 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4321 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4441 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4501 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4561 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4621 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4681 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4741 ggagcagaca agcccgtcag ggcgcgtcag cgggt
//
