LOCUS       sequence-210329-        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_670 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5108)
  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 5108)
  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_670
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          132..228
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(289..291,292..294,295..1005)
                     /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(334..355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(595..614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     942..963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1707..1841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1731..1750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1785..1804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1952..1968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1952..1968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1965..1987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1976..1992
                     /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(2000..2006,2007..2024,2025..2030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2045..2066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2183..2200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2354..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2434..2453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3113..3904,3905..3973))
                     /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     3736..3755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3974..4078)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4146..4164
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4202..4224)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4324..4343
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4537..4559
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4551..4568
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4552..4568
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4595..4898
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4899..5102
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5053..5073
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5099..5108,1..15)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
       61 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      121 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      181 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      241 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag
      301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      361 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      481 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      541 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      721 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
      781 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      841 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
      901 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      961 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgaat tggtggaggg
     1021 ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga gatctgatat gtctgacacc
     1081 ccagcgggaa aactcccggc tgggatgttt ctccgtttat cgaatttttc caggagaaac
     1141 atcatgtcta ctacatctaa aaccaccacc gctaaaccta aaaaaagcag tgccagcaag
     1201 aaagccccgg ctccagttcc tgctgaatca gtaaattcgg ttcagtcagc agctccggca
     1261 caaccaaaac cacaatccag ctacttcaat ttgcatatcg atggactggg ttacctgagc
     1321 aatattcgcg aaattaatac tgctacgggt tcattcctga gctgtacgat taacgcgtta
     1381 cacggctcaa ctgaaaaacc tgaatacacg cgttttgatg tcacggtgtc gggtaaacag
     1441 accacttctt taatcagacg ctgccagaat gcagtgaatg aagagaaaaa ggtacttatc
     1501 ggttttaagc tcagtaatct gactgccgat gtgtttactt taaccaaagg cgatcatgcg
     1561 ggtgaatcaa gagcgtcact gaaagcgaga ctgataaaaa tcagttttat caaagtgggg
     1621 cagaatatgg tttataaagc tgaaaagtcc gagtcatccg actcatcata ctgaatcggt
     1681 accctcgaga agcttgcggc cgcggggatc cagacatgat aagatacatt gatgagtttg
     1741 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1801 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1861 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct ctagagtcga
     1921 tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2041 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2101 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2761 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctgtgc
     2821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4081 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4141 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4201 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4261 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4321 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     4381 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     4441 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4501 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4561 cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
     4621 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4681 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4741 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4801 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4861 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     4921 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4981 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5041 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5101 ctcgttta
//
