LOCUS       sequence-213511-        5492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_052 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5492)
  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 5492)
  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_052
FEATURES             Location/Qualifiers
     source          1..5492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..27)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     355..376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1504..1638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1528..1547)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1582..1601
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1749..1765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1749..1765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1762..1784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1773..1789
                     /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(1797..1803,1804..1821,1822..1827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1842..1863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1980..1997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2151..2739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2231..2250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2910..3701,3702..3770))
                     /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     3533..3552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3771..3875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3943..3961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3999..4021)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4121..4140
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4334..4356
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4348..4365
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4349..4365
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4392..4695
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4696..4899
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4850..4870
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4896..4920
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5037..5133
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(5194..5196,5197..5199,5200..5492,1..418)
                     /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(5239..5260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
       61 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      121 caagctggag tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa
      181 cggcatcaag gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc
      241 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      301 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
      361 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagcg
      421 aattggtgga gggggcggtt ccggtggcgg aggttcccga attcgtcgac ggagatctga
      481 tatgcaaaaa cgttatctgg gtaaatccgg actcgaagtc tccgcccttg ggctcggttg
      541 catgggctta agccacggct acggcccggc gaccgatacc cgtcaggcta tcgaacttat
      601 tcgtgctgcg gttgaacgtg gcgtcacctt cttcgatacc gccgaagtgt atggtccgta
      661 tctgaatgaa gaagtggtcg gcgaagcctt aaaaccgttt cgtgaccgcg tggtaattgc
      721 cactaaattc ggttttacct tcggcaatga taataagcag cagattttaa acagccgtcc
      781 ggaacatatc cgtgaagccg ttgaaggatc attacgtcgt ctgaaaacag atgttattga
      841 cctgctgtat caacaccgtg ttgatccaga tgttcctatt gaagatgttg caggaacggt
      901 gaaagacctc atcgcagaag gtaaggtcaa acatttcggc ctgtctgaag caggagcaca
      961 aaccattcgt cgtgcacatg cggtacaacc tgtcaccgcc ctgcaaagcg aatattccat
     1021 gtggtggcgt gagcctgaac aggaaattct gccgttgctt gaagaactgg gaattggttt
     1081 tgtccccttc agcccactag gtaaaggttt cctgacaggg gcgattaagc cgggaaccac
     1141 tttcggcaaa gatgattacc gcagcaccgt accgcgtttc gcagcacaag ccattgaagc
     1201 caatgaaaaa ctggtcacct tgttaggtga actggctgcg gagaaagggg taacgtctgc
     1261 acaaattgca ctggcatggc tattagcaca aaagccgtgg attgttccca tccctggcac
     1321 cactaaactg caccggctgg aagaaaacct ggccgctgct gacattgttc tttcgcagaa
     1381 agatacgcag cagataagcg aggcgctgga aacaattaaa atcgtcggtg aacgttactc
     1441 gcctgaacat caggcacgcg taggccgctg aatcggtacc ctcgagaagc ttgcggccgc
     1501 ggggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1561 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1621 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1681 gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg
     1741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2341 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2581 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3301 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3901 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3961 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4021 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4081 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4141 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4201 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4261 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     4321 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca
     4381 tgcctgcagg tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4441 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4501 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4561 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4621 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4681 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4741 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4801 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4861 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     4921 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
     4981 ctccggactc tagaggatcc ggtactagag gaactgaaaa accagaaagt taactggtaa
     5041 gtttagtctt tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa
     5101 ctgctcctca gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc
     5161 taaaagctgc ggaattgtac ccgcgggccc accatggtga gcaagggcga ggagctgttc
     5221 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5281 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     5341 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg
     5401 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     5461 cccgaaggct acgtccagga gcgcaccatc tt
//
