LOCUS       sequence-210449-        5267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_678 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5267)
  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 5267)
  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_678
FEATURES             Location/Qualifiers
     source          1..5267
                     /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    1863..1997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1887..1906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1941..1960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2108..2124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2108..2124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2121..2143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2132..2148
                     /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(2156..2162,2163..2180,2181..2186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2201..2222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2339..2356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2510..3098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2590..2609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3269..4060,4061..4129))
                     /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     3892..3911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4130..4234)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4302..4320
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4358..4380)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4480..4499
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4693..4715
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4707..4724
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4708..4724
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4751..5054
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5055..5258
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5209..5229
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5255..5267,1..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 ctgatatgtc gcagatgaac
     1081 acgtcttcac ccccggtaca gtcgtcaacg ccaccaaagc cgcacggttt ttttaccgta
     1141 ctgctgtggg atacgccgat gaacattatc ggcatgctgc tggtttcgtt attattcagt
     1201 gtaattgtgg agttcatcgg tattgcgttc ttttggcctg atcagggagc ggaacatagc
     1261 cgcgccgtta tgctgacgga gagcggttat tttgccgaag gatttacgca aagtttgctg
     1321 ctgtcatcgc cggtggtgac cgtacagagc tggatcgcgc tgggttatca gtggtttttt
     1381 gtgaatagcg gttttactgg ctggttgcag tcgcttcaga cgacgactgc cggaggcggt
     1441 aatggcgtta cggatgaact taacgcctgg tgtgcctggg cagtacagat tgcatatgaa
     1501 tacctgatgg cgacgatgta tgtgtcgctg gtcttcctga tgcgggtcac cattctggtg
     1561 ctgtcggtgc cgctatttgt tctggttgcc attgtgggga tcgcggatgg gatggttcga
     1621 cgcgatctcc gtcgctatgg tgctgcgtat gagtcgagct tcctctacca tcatgccaaa
     1681 cgcgtggtga agccagcagc atatattccg tgcgtattgt atttgtcagc gccgttcgcg
     1741 gtatatccga atttgttgct gttaccggca gcattgctga tggggctggc gatttcagtg
     1801 acgatgggga gttttaagaa gtatttgtga atcggtaccc tcgagaagct tgcggccgcg
     1861 gggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1921 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1981 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2041 aggtgtggga ggttttttcg gatcctctag agtcgatctg caggcatgct agcttggcgt
     2101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4261 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4321 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4381 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4441 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4501 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4561 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4621 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     4681 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagttc gagcttgcat
     4741 gcctgcaggt cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4801 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4861 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4921 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4981 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     5041 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     5101 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5161 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5221 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtg
//
