LOCUS       sequence-212754-        4901 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_810 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4901)
  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 4901)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_810
FEATURES             Location/Qualifiers
     source          1..4901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..138
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     130..147
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     131..147
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        174..477
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        478..681
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     632..652
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     678..702
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          819..915
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(976..978,979..981,982..1692)
                     /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(1021..1042)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1282..1301)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1629..1650
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2187..2321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2211..2230)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2265..2284
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2432..2448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2432..2448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2445..2467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2456..2472
                     /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(2480..2486,2487..2504,2505..2510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2525..2546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2663..2680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2834..3422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2914..2933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3593..4384,4385..4453))
                     /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     4216..4235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4454..4558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4626..4644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4682..4704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4804..4823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
       61 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      121 tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca
      181 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      241 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      301 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      361 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      421 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      481 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      541 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      601 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      661 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
      721 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat
      781 ccggtactag aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt
      841 ttatttcagg tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt
      901 tgcctttact tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt
      961 acccgcgggc ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1021 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1081 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1141 gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt cgcccgctac
     1201 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1261 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1321 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1381 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta tatcaccgcc
     1441 gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat cgaggacggc
     1501 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1561 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     1621 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1681 gagctgtaca agcgaattgg tggagggggc ggttccggtg gcggaggttc ccgaattcgt
     1741 cgacggagat ctgatatgaa cctaagcggg caatcattag tttggacttg cttcaatgct
     1801 ctgtgcctgg aagattttca tctgaatgct ttgtgggaga aattttcaat cctgagaata
     1861 gactttccgg ttgttagttt ggctcatcgg gatatgtttt cgtttttacc caataaaaag
     1921 gtcttactga tgaaagagaa tattcctgct gttacgcgta cacgtattga tgttggctgc
     1981 tgtgacagca gtgcagtttt tgtgcttgat aatggtcacg atcatttcgc tgtttccctt
     2041 gaaacggtac tctcatgtct gaaggtggca gaggaacacg gccatgtacc tgaaattggc
     2101 gcggattggt ggtgtcttat cgctaaccat actgcgctgg gatacccgtc atgaatcggt
     2161 accctcgaga agcttgcggc cgcggggatc cagacatgat aagatacatt gatgagtttg
     2221 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2281 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2341 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct ctagagtcga
     2401 tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2461 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2521 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4561 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4621 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4801 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     4861 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc a
//
