LOCUS       sequence-210528-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_776 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  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 4952)
  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_776
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          128..224
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(285..287,288..290,291..1001)
                     /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(330..351)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(591..610)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     938..959
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1547..1681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1571..1590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1625..1644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1792..1808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1792..1808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1805..1827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1816..1832
                     /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(1840..1846,1847..1864,1865..1870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1885..1906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2023..2040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2194..2782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2274..2293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2953..3744,3745..3813))
                     /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     3576..3595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3814..3918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3986..4004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4042..4064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4164..4183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4377..4399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4391..4408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4392..4408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4435..4738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4739..4942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4893..4913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4939..4952,1..11)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
       61 accgatccag cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag
      121 ttaactggta agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc
      181 aaatcaaaga actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt
      241 tacttctgct ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg
      301 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      361 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      421 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga
      481 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      541 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      601 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      661 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      721 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact
      781 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      841 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      901 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      961 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gcgaattggt ggagggggcg
     1021 gttccggtgg cggaggttcc cgaattcgtc gacggagatc tgatatggag atcgtgtacg
     1081 atcccgtcat atgtaaggaa caaggtgtca ataacttcgc gcgtctggcc gacgaaatgg
     1141 caggcattat tcaacgcgat gagtgccgtc tgggaattta catttgtggg actggcattg
     1201 gctttacttg tcagatcaac aaacactggg gcataagagc cgttgcggtt actaacccct
     1261 actccgcaaa gcgggcgcga ctcagcaata acgcacaagt tattggactg ggttgccggg
     1321 tcaacgatct cgaatacagc aaaatgattg tcgatgcatg gttggataac ccattcgact
     1381 tcaccaccgc cagagaaaat tctaaaaaga acttactgga agctgaacgt aacgataatg
     1441 cattgctggt taagcccgct gatgtacgct ggaatatggg tttccgacca gatgatgaaa
     1501 aatctgaggg ttgaatcggt accctcgaga agcttgcggc cgcggggatc cagacatgat
     1561 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1621 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1681 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1741 ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat ggtcatagct
     1801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3961 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4021 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4081 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4141 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4201 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     4261 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4321 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     4381 gtcacgacgt tgtaaaacga cggccagtga gttcgagctt gcatgcctgc aggtcgttac
     4441 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     4501 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     4561 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     4621 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     4681 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4741 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     4801 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4861 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     4921 ggaggtctat ataagcagag ctcgtttagt ga
//
