LOCUS       sequence-212759-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_762 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  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 4682)
  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_762
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..132
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     124..141
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     125..141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        168..471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        472..675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     626..646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     672..696
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          813..909
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(970..972,973..975,976..1686)
                     /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(1015..1036)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1276..1295)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1623..1644
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1962..2096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1986..2005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2040..2059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2207..2223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2207..2223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2220..2242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2231..2247
                     /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(2255..2261,2262..2279,2280..2285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2300..2321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2438..2455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2609..3197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2689..2708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3368..4159,4160..4228))
                     /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     3991..4010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4229..4333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4401..4419
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4457..4479)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4579..4598
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
       61 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
      121 cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc tgcaggtcgt tacataactt
      181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      241 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      301 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      361 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      421 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      481 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      541 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      601 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      661 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
      721 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctaga ggatccggta
      781 ctagaggaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg tcttttattt
      841 caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt
      901 tacttctagg cctgtacgga agtgttactt ctgctctaaa agctgcggaa ttgtacccgc
      961 gggcccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1021 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1081 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1141 tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt gcttcgcccg ctaccccgac
     1201 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1261 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1321 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1381 ctggggcaca agctggagta caactacaac agccacaagg tctatatcac cgccgacaag
     1441 cagaagaacg gcatcaaggt gaacttcaag acccgccaca acatcgagga cggcagcgtg
     1501 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1561 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1621 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1681 tacaagcgaa ttggtggagg gggcggttcc ggtggcggag gttcccgaat tcgtcgacgg
     1741 agatctgata tgagccgcgt aatcaatttg ggtaaggaga agaaattccc aattactcaa
     1801 gagctatacg agcggcttga aagcgtcatc catgattacg atggtgaaat aagtttatgc
     1861 gaggcgattg gcacactcga attgctgaag cagtcactga ttgaaggcga gaaagagtcc
     1921 tcaacctgaa tcggtaccct cgagaagctt gcggccgcgg ggatccagac atgataagat
     1981 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2041 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2101 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcgg
     2161 atcctctaga gtcgatctgc aggcatgcta gcttggcgta atcatggtca tagctgtttc
     2221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2281 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2521 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4381 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4441 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4501 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4561 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     4621 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     4681 gg
//
