LOCUS       sequence-212784-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_441 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  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 4946)
  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_441
FEATURES             Location/Qualifiers
     source          1..4946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     129..146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     130..146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        173..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          818..914
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(975..977,978..980,981..1691)
                     /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(1020..1041)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1281..1300)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1628..1649
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2231..2365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2255..2274)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2309..2328
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2476..2492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2476..2492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2489..2511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2500..2516
                     /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(2524..2530,2531..2548,2549..2554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2569..2590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2707..2724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2878..3466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2958..2977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3637..4428,4429..4497))
                     /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     4260..4279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4498..4602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4670..4688
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4726..4748)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4848..4867
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
       61 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      121 cacgacgttg taaaacgacg gccagtgagt tcgagcttgc atgcctgcag gtcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagaggatc
      781 cggtactaga ggaactgaaa aaccagaaag ttaactggta agtttagtct ttttgtcttt
      841 tatttcaggt cccggatccg gtggtggtgc aaatcaaaga actgctcctc agtggatgtt
      901 gcctttactt ctaggcctgt acggaagtgt tacttctgct ctaaaagctg cggaattgta
      961 cccgcgggcc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1021 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1081 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1141 tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc
     1201 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1261 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1321 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1381 acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg
     1441 acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca
     1501 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1561 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     1621 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1681 agctgtacaa gcgaattggt ggagggggcg gttccggtgg cggaggttcc cgaattcgtc
     1741 gacggagatc tgatatgact ataaatatat taaaagaaaa gctaaagtta acttgcggag
     1801 ccatatttat atttttttta tccacgactt ttgttttctt tggctgttat gctacttatg
     1861 aattattttt atggaatgat attattgtat atagctgggg atatatatta attgtctttt
     1921 tacctttcac attatatgta atgtcgtttg agattttgtt ttttgctatt agtgggcgac
     1981 gattgtctaa agtaacaatg gtgcgccttt ggttgataat taaaattatt attgctttct
     2041 ctatttgcgc agtgttgatt ttttcttcaa tttacaaaaa agaattatta tctagaaatt
     2101 atattgcttg tagtggtatc ccgtctgggt ggatgccggg tctggcaacg aaatacgtta
     2161 aagaaaaatc attatgcgaa aaaaatggca ataattgaat cggtaccctc gagaagcttg
     2221 cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2281 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2341 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2401 tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgatctgca ggcatgctag
     2461 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2521 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2581 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2641 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2701 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2761 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2821 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2881 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2941 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3001 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3061 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3121 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3181 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3241 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3301 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3361 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3421 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3481 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3541 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3601 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3661 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3721 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3781 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3841 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3901 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3961 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4021 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4081 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4141 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4201 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4261 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4321 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4381 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4441 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4501 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4561 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4621 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4681 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4741 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4801 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4861 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4921 ataccgcatc aggcgccatt cgccat
//
