LOCUS       sequence-210157-        5162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_352 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5162)
  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 5162)
  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_352
FEATURES             Location/Qualifiers
     source          1..5162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..70
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     62..79
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     63..79
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        410..613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     610..634
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          751..847
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(908..910,911..913,914..1624)
                     /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(953..974)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1214..1233)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1561..1582
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2380..2514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2404..2423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2458..2477
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2625..2641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2625..2641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2638..2660)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2649..2665
                     /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(2673..2679,2680..2697,2698..2703))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2718..2739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2856..2873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3027..3615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3107..3126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3786..4577,4578..4646))
                     /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     4409..4428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4647..4751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4819..4837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4875..4897)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4997..5016
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
       61 ttgtaaaacg acggccagtg agttcgagct tgcatgcctg caggtcgtta cataacttac
      121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      601 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
      661 tgacctccat agaagacacc gggaccgatc cagcctccgg actctagagg atccggtact
      721 agaggaactg aaaaaccaga aagttaactg gtaagtttag tctttttgtc ttttatttca
      781 ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat gttgccttta
      841 cttctaggcc tgtacggaag tgttacttct gctctaaaag ctgcggaatt gtacccgcgg
      901 gcccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc ttcgcccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacaacag ccacaaggtc tatatcaccg ccgacaagca
     1381 gaagaacggc atcaaggtga acttcaagac ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caagcgaatt ggtggagggg gcggttccgg tggcggaggt tcccgaattc gtcgacggag
     1681 atctgatatg ggtgataatt ataatattat accgcaaaag aaattcacat tatctggtgt
     1741 ttataggata aagaatgcag agagttcaat agagcttgcc attcgttcaa ttatagatgt
     1801 aatggatgaa gtgattgttg tagataatga atcttcagat ggtacattag atattttaat
     1861 taaactgcaa aaaaaatatc ctaataaaat aaagattttc cattataaca aaaaactgtg
     1921 tagagcaggt aaaaattatg ctgattgtgt gagaagtaat ccatcaggga gcttagccaa
     1981 atattataac tatgcttttt ccaaagcaac tagcgaatat gtaatgaagt gtgatgccaa
     2041 ttatattttc acgttgaaag gtaaaataga tattatcaat gcattaaata agaatcctga
     2101 tgtattatgc tatcctggtg ttgaaatatt tgggcatcat cactcaattg agccatttgt
     2161 atatttaagg aaattaaatt ataaatactg tgatggtttg ttatgggagt ttttacatta
     2221 tgaacgaact gcgaaaataa agaaaatatt aaatccgtgc tttgtgcata taaaaagaat
     2281 caattacaat aagtatatac atagcgaaca taaaggagtt gacggtttat atggtagcca
     2341 cgattgaatc ggtaccctcg agaagcttgc ggccgcgggg atccagacat gataagatac
     2401 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2461 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2521 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttcggat
     2581 cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata gctgtttcct
     2641 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2701 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2761 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2821 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2881 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2941 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3001 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3061 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3121 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3181 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3241 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3301 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3361 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3421 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3481 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3541 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3601 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3661 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3721 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3781 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3841 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3901 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3961 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4021 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4081 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4141 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4201 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4261 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4321 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4381 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4441 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4501 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4561 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4621 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4681 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4741 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4801 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4861 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4921 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4981 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5041 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     5101 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     5161 aa
//
