LOCUS       sequence-210543-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_240 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  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 5513)
  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_240
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          130..226
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(287..289,290..292,293..1003)
                     /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(593..612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     940..961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2110..2244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2134..2153)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2188..2207
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2355..2371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2355..2371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2368..2390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2379..2395
                     /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(2403..2409,2410..2427,2428..2433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2448..2469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2757..3345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2837..2856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3516..4307,4308..4376))
                     /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     4139..4158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4377..4481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4549..4567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4605..4627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4727..4746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4940..4962
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4954..4971
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4955..4971
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4998..5301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5302..5505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5456..5476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5502..5513,1..13)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
       61 ggaccgatcc agcctccgga ctctagagga tccggtacta gaggaactga aaaaccagaa
      121 agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt
      181 gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt
      241 gttacttctg ctctaaaagc tgcggaattg tacccgcggg cccaccatgg tgagcaaggg
      301 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      361 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      421 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
      481 gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
      541 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      601 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      661 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      721 ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa
      781 cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      841 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      901 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      961 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcgaattg gtggaggggg
     1021 cggttccggt ggcggaggtt cccgaattcg tcgacggaga tctgatatga gcagtaaagt
     1081 cagtggcatg aatacgttgc tctccgttag cgtgcggact aataaccctg aacctggttt
     1141 ccagttgatt aaccagcgaa tcacccacag cacgattaac gacaatattt gggcatcgct
     1201 acaaaatgct caaattcagg cgttgaaaac gctcgaccag cgtccagcgg agaaattcgc
     1261 ccagcagatg tatgagacgc gctatgccga tgaccgcacg aaattaccgc aagaaaatca
     1321 gattgcacag tttactgccg cagatgcgct ggctgccgat cgccagttgt tttcatctcc
     1381 agcggatatc acgtttgtca ttgtcggtaa tgtcgcagaa gacaaactcg tggcgttaat
     1441 tacgcgttac ttaggatcta tcaaacactc tgattcgcca ttagccgcag gtaaaccatt
     1501 aactcgcgcg acggacaacg catcggttac tgtaaaagaa caaaatgaac cagtggcaca
     1561 ggtttcacag tggaagcgtt ataattcccg gacacctgtt aatctggcga cgcgtatggc
     1621 gctcgatgct tttaacgtcg cactggcaaa agatctacgc attaatattc gtgaacaggc
     1681 atctggggca tacagcgttt cttctcgcct ctcggttgat cctcaggcca aagatatcag
     1741 tcatttgctg gcttttactt gtcaaccaga acgacatgat gaactgttaa cgttagcgaa
     1801 tgaagtgatg gttaagcgtc tggccaaagg gatcagtgag caagaactga acgaatacca
     1861 gcaaaacgtt cagcgcagcc tcgatatcca acagcgtagc gttcaacaat tagcgaacac
     1921 cattgtaaat agtcttattc aatatgacga tcctgcagca tggattgagc aggagcaatt
     1981 gttgaaacaa atgacggtag agaatgttaa cactgctgtt aaacaatatc tttctcatcc
     2041 ggtgaatacg tataccggag tattattgcc aaaatgaatc ggtaccctcg agaagcttgc
     2101 ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2161 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2221 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2281 cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag gcatgctagc
     2341 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4501 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4561 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4621 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4681 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4741 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4801 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4861 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     4921 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagttcgag
     4981 cttgcatgcc tgcaggtcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     5041 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     5101 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     5161 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     5221 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     5281 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     5341 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     5401 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     5461 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agt
//
