LOCUS       sequence-210463-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_700 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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_700
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          129..225
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(286..288,289..291,292..1002)
                     /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(331..352)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(592..611)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     939..960
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2547..2681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2625..2644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2792..2808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2792..2808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2805..2827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2816..2832
                     /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(2840..2846,2847..2864,2865..2870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2885..2906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3023..3040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3194..3782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3274..3293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3953..4744,4745..4813))
                     /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     4576..4595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4814..4918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4986..5004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5042..5064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5164..5183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5377..5399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5391..5408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5392..5408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5435..5738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5739..5942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5893..5913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5939..5951,1..12)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
       61 gaccgatcca gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa
      121 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
      181 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
      241 ttacttctgc tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc
      301 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      361 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      421 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg
      481 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      541 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      601 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      661 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      721 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
      781 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      841 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      901 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      961 accgccgccg ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc
     1021 ggttccggtg gcggaggttc ccgaattcgt cgacggagat ctgatatgaa agagaaaaaa
     1081 acatccctca ccaataatga gaatgaattc gatataactg ttttcccctc aattgtggaa
     1141 aagagcaaat accaagactc actgctcgcc agttccaccg acgccagcaa ggctctagag
     1201 gcactgacga agaactctgg cgtggatgca ctcctcaaaa accatgattc actgtttgcc
     1261 ggttccactg ccgccagcaa ggcattcgag gcaatgacac gcaactccgt tgcgaacact
     1321 ttcctcaaaa accatgattc actgcttgcc agctccactg ccgccagcaa ggcattcgag
     1381 gcaatgacac gcaactccgt tgcgaacact ttcctcaaaa accatgattc actgcttgcc
     1441 agctccactg ccgccagcaa ggcattcgag gcaatgacac gcaactccgt tgcgaacact
     1501 ttcctcaaaa accatgattc actgtttgcc ggttccactg ccgccagcaa ggcattcgag
     1561 gcaatgacac gcaactccgt tgtgaactct ttcctcaaaa accatgattc actgtttgcc
     1621 ggttccactg ccgccagcaa ggcattcgag gcaatgacac gcaactccgt tgtgaactct
     1681 ttcctcaaaa accatgattc actgtttgcc agctccactg ccgccagcaa ggcattcgag
     1741 gcaatgacac gcaactccgt tgtgaactct ttcctcaaaa accatgattc actgtttgcc
     1801 agctccactg ccgccagcaa ggcattcgag gcaatgacgc ggaactctgg tgtggccact
     1861 cttttcgcaa agcaaagaga atcatttacc cgggattatt ttactacgag tattactcaa
     1921 cacttagata aattaaatcc aaccgagatt caaaaattac aggaagctgc tattctattt
     1981 tcgcgctcac ctcaaggcgc agccgttatt gcaaccaata tcaatacagt agaaataaca
     2041 gagcccgacg cattaacaaa attaaaaaaa tacatcgaag aaagccctgt cacttcacat
     2101 tttaggaaat tacctttatt agtacagctt atcataattt atcttgtaat gactatgtcc
     2161 tctgttattt ccgataaaaa caaagagtta gcaatttatt taattgaaga atcacaacaa
     2221 tattttagca aagtagaatc accaaacgcc catgttaaga aattgacaaa acagttaccc
     2281 aatgaaattg atgtaagtac attaaaacat atcagaataa ttactggcga aaacgtccga
     2341 ttacggaata gaccatcaat gcatggtgat gtattggtca cgttgcaaaa atacacacct
     2401 gttatcgtta tcgataaaag tgatagaaaa tggctatata tacaactatc acttggagat
     2461 caaaagatct atggctgggt caatcgttcc tataccaaag cattaaatca ctgaatcggt
     2521 accctcgaga agcttgcggc cgcggggatc cagacatgat aagatacatt gatgagtttg
     2581 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2641 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2701 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct ctagagtcga
     2761 tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2821 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2881 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2941 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3001 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3061 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5161 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5221 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     5281 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5341 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5401 cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
     5461 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5521 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5581 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5641 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5701 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     5761 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5821 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5881 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5941 ctcgtttagt g
//
