LOCUS       sequence-213513-        4955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_252 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4955)
  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 4955)
  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_252
FEATURES             Location/Qualifiers
     source          1..4955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..131
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     123..140
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     124..140
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        167..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..695
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          812..908
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(969..971,972..974,975..1685)
                     /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(1014..1035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1275..1294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1622..1643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2234..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2258..2277)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2312..2331
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2479..2495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2479..2495)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2492..2514)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2503..2519
                     /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(2527..2533,2534..2551,2552..2557))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2572..2593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2710..2727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2881..3469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2961..2980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3640..4431,4432..4500))
                     /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     4263..4282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4501..4605)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4673..4691
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4729..4751)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4851..4870
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
       61 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      121 gttgtaaaac gacggccagt gagttcgagc ttgcatgcct gcaggtcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
      721 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag gatccggtac
      781 tagaggaact gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc
      841 aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt
      901 acttctaggc ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg
      961 ggcccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1021 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1081 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1141 ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc
     1201 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1261 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1321 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1381 tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc gccgacaagc
     1441 agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac ggcagcgtgc
     1501 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1561 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1621 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1681 acaagcgaat tggtggaggg ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga
     1741 gatctgatat ggataaacct gcgaatcgta ttgaagatat ccacggtatg gctgatttca
     1801 tcagccagta tcctggtgtt gatagcaccc gtattggttt gttggggatt tgcgggggag
     1861 gagggtattc acttgctgcc gccgagacgg acaaacgatt caaatcgatt gcaaccatta
     1921 gcatgtttaa ttcaggactt gtacggcgca atggtatgca ggattcacag ttggatacta
     1981 ttcaacaacg tcttaagcag gcttcagatg cacgagctca ggaagtcgcc ggtagcgaag
     2041 tgctttactc cggtgatgcc aacctgactg atgaacagat cgctaaatta ccttttgcgt
     2101 tatatcgcca gggctacgag tactactgga aaacccacgc acacccaaac atattcagaa
     2161 gcgtgaggga tatagtgcca tcaaagcgct ggcagctgtg aatcggtacc ctcgagaagc
     2221 ttgcggccgc ggggatccag acatgataag atacattgat gagtttggac aaaccacaac
     2281 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2341 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2401 ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc
     2461 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2521 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2581 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3061 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3301 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3481 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3541 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4621 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4681 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4741 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4801 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4861 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4921 aaaataccgc atcaggcgcc attcgccatt caggc
//
