LOCUS       sequence-212783-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_136 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  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 5891)
  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_136
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     183..205
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     197..214
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     198..214
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        241..544
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        545..748
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     699..719
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     745..769
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          886..982
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1043..1045,1046..1048,1049..1759)
                     /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(1088..1109)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1349..1368)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1696..1717
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3244..3378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3268..3287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3322..3341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3489..3505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3489..3505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3502..3524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3513..3529
                     /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(3537..3543,3544..3561,3562..3567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3582..3603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3720..3737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3891..4479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3971..3990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4650..5441,5442..5510))
                     /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     5273..5292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5511..5615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5683..5701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5739..5761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5861..5880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
       61 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      121 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      181 ttcccagtca cgacgttgta aaacgacggc cagtgagttc gagcttgcat gcctgcaggt
      241 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      301 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      361 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      421 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      481 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      541 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      601 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      661 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      721 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      781 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      841 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      901 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      961 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
     1021 gaattgtacc cgcgggccca ccatggtgag caagggcgag gagctgttca ccggggtggt
     1081 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1141 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1201 gctgcccgtg ccctggccca ccctcgtgac caccttgacc tacggcgtgc agtgcttcgc
     1261 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1321 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1381 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1441 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca aggtctatat
     1501 caccgccgac aagcagaaga acggcatcaa ggtgaacttc aagacccgcc acaacatcga
     1561 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1621 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     1681 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1741 catggacgag ctgtacaagc gaattggtgg agggggcggt tccggtggcg gaggttcccg
     1801 aattcgtcga cggagatctg atatggctac ttggcaagga tcaaatggcg gattgttagc
     1861 tggtatcggc ggcgtcaact caaacgctcc gagcgtaaat gacatcggca atacgcttca
     1921 gcttatcagg cagaacaatg atattgagcg ttcaggcgct aacaatgctg ggctgactgc
     1981 tttgcaaggc ctttcaggta ttgcgggggt gtttcagcag gagaagcagg ctcagcggca
     2041 gaaaaaattt cagcaggcgt acgctaatgc ttatgcgtct ggtgatcgcg gtgctttgcg
     2101 tcagttggct actcaatatc cagaccagat tgaatcagtt cgtaaaggca tgggattcat
     2161 tgatgaagac cagcgcaatt ctatcggcac attagcggct ggcgcacgcc ttgcgtcatc
     2221 ttctccagaa gcaatgcaat catggctgca aaacaacgcc aaggaactga ctcgcgtcgg
     2281 tgttgaccct aacagcgttg ctcagatgta tcagcagaac ccttcaggat ttggtgagtt
     2341 tgttgatcac cttggaatgg ctgctcttgg tccgattgat tacttcaatg ttcaggacaa
     2401 gatggctggt cgtgagattg atcgcggaaa acttgcagag acaatccgca gcaatcaggc
     2461 tggcgaggcg ctaacagcac gaggccagaa catcacgatg cgcggtcagg acttatctgc
     2521 ttctactgcg cgacgcgggc aggatttggc aatgcagcga gcgtcaacaa gaggaaccgc
     2581 tgggaatgat gagcgtacag ttcagttatc agatggcaga actgtaacgg taggtgggaa
     2641 gcttcacggc gctggggcca atgcgttcta cgaaggcatc gacaacgagg ggaatatggt
     2701 tcgcgtccct gccagttcaa tcgctgcgcc tgcaacatcg tctgcatcag cacaaaacta
     2761 tgccatgaag aaggatatcg acgcgatcgc aaatgcagac gcttctgctc tcgatttcat
     2821 gacaggaatg acaggcggtg caggtaatcc agcaattggt gctgatgttc gcagcagatt
     2881 aacaggaaaa gagcagcgcc agttatataa ctcagcacaa cggattcagg gcagaatgca
     2941 gaatcagggt gtggcggcag caagggacat gggtgccagt ggtattaaca ccgttgcaga
     3001 agcgaagatg tattttcagg ggatgccgca ggttgactat tcaagcccgg aggctatgca
     3061 gcagtcgatt cgtgagattc aggaatacac caacaattac aaccaacaat ataacgttaa
     3121 tgttggtaaa tctcagcggc agcaatctca acctgcacag gtatcacagc cagcagccag
     3181 cagtaacttt tcttcactat ggggtgattg aatcggtacc ctcgagaagc ttgcggccgc
     3241 ggggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3301 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3361 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3421 gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg
     3481 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3541 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3601 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3661 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3721 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3781 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3841 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3901 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3961 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4021 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4081 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4141 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4201 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4261 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4321 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4381 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4441 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4501 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4561 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4621 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4681 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4741 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4801 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4861 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4921 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4981 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5041 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5101 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5161 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5221 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5281 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5341 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5401 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5461 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5521 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5581 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5641 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5701 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5761 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5821 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5881 ctgagagtgc a
//
