LOCUS       sequence-210537-        4706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_808 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4706)
  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 4706)
  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_808
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          130..226
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(287..289,290..292,293..1003)
                     /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(332..353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(593..612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     940..961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1303..1437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1327..1346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1381..1400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1548..1564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1548..1564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1561..1583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1572..1588
                     /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(1596..1602,1603..1620,1621..1626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1641..1662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1779..1796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1950..2538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2030..2049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2709..3500,3501..3569))
                     /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     3332..3351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3570..3674)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3742..3760
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3798..3820)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3920..3939
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4133..4155
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4147..4164
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4148..4164
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4191..4494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4495..4698
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4649..4669
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4695..4706,1..13)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
       61 ggaccgatcc agcctccgga ctctagagga tccggtacta gaggaactga aaaaccagaa
      121 agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt
      181 gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt
      241 gttacttctg ctctaaaagc tgcggaattg tacccgcggg cccaccatgg tgagcaaggg
      301 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      361 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      421 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
      481 gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
      541 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      601 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      661 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      721 ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa
      781 cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      841 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      901 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      961 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcgaattg gtggaggggg
     1021 cggttccggt ggcggaggtt cccgaattcg tcgacggaga tctgatatga catcgaaata
     1081 cacttatctt ccggttgcgg atgaacgagc ttcctggcgt agtcagtcca ttatggcgct
     1141 ggaaataaca gcggatatca actgtaagcg tgcaactgaa cgtgaccggc aaaatttcct
     1201 ttctgccagg aagcgtttgc aggaacgtgt taacagcgtg ctggcttccg gggaggtatg
     1261 tcatggatga atcggtaccc tcgagaagct tgcggccgcg gggatccaga catgataaga
     1321 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1381 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1441 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttcg
     1501 gatcctctag agtcgatctg caggcatgct agcttggcgt aatcatggtc atagctgttt
     1561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     3721 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     3781 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     3841 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     3901 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     3961 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     4021 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4081 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4141 cgacgttgta aaacgacggc cagtgagttc gagcttgcat gcctgcaggt cgttacataa
     4201 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4261 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4321 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4381 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     4441 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4501 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4561 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4621 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4681 gtctatataa gcagagctcg tttagt
//
