LOCUS       sequence-210478-        5135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_723 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5135)
  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 5135)
  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_723
FEATURES             Location/Qualifiers
     source          1..5135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          135..231
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(292..294,295..297,298..1008)
                     /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(337..358)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(598..617)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     945..966
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1737..1871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1761..1780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1815..1834
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1982..1998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..1998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1995..2017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2006..2022
                     /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(2030..2036,2037..2054,2055..2060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2075..2096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2213..2230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2384..2972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2464..2483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3143..3934,3935..4003))
                     /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     3766..3785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4004..4108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4176..4194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4232..4254)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4354..4373
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4567..4589
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4581..4598
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4582..4598
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4625..4928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4929..5132
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5083..5103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5129..5135,1..18)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
       61 caccgggacc gatccagcct ccggactcta gaggatccgg tactagagga actgaaaaac
      121 cagaaagtta actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg
      181 gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg
      241 gaagtgttac ttctgctcta aaagctgcgg aattgtaccc gcgggcccac catggtgagc
      301 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      361 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      421 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      481 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      541 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      601 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      661 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      721 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
      781 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
      841 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      901 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      961 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagcg aattggtgga
     1021 gggggcggtt ccggtggcgg aggttcccga attcgtcgac ggagatctga tatgaaaatt
     1081 caaaacaaac atgttattgc ctggctggag agttgtgctg cacatcttac tgaacaacaa
     1141 gattttctaa ccgcacttga cagagatatt ggtgatgccg accacggcct caatatgaat
     1201 cgtggtttca gtgccgtgaa agcgacgttg cccgacattg aacgtcagca tattggcaat
     1261 atccttaaaa atactggcat gaaactactt tccagcgttg gcggtgccag tggtcctctt
     1321 tacggcacat tatttattcg cgcttctgcg gcagtgggag ccagaacaga attaagtctg
     1381 gaagagtggc tggcgtgtct ggaagaaggg attgcaggcg ttatcgcacg aggaaaagca
     1441 gaacaagggg ataaaacact gtgtgatgtc tgggttccag tcctccacga agccaaaaag
     1501 aacctgcaag ccggaatgtc gccttccctt ctgctgaata cgatggtgca agatgcggca
     1561 acagccgtcg ataacaccat caatatgcag gcgaaaaaag gacgtgccag ttatctcggc
     1621 gctcgcagtg tgggccatca ggatccaggc gcgacctcct cctggctgat gattaaggct
     1681 atgcaggaag gattagcggg atgaatcggt accctcgaga agcttgcggc cgcggggatc
     1741 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1801 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1861 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1921 gggaggtttt ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat
     1981 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2041 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4141 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4201 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4261 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4321 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4381 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     4441 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4501 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     4561 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gttcgagctt gcatgcctgc
     4621 aggtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4681 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4741 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4801 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4861 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4921 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4981 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     5041 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     5101 gtgtacggtg ggaggtctat ataagcagag ctcgt
//
