LOCUS       sequence-210505-        5390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_730 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  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 5390)
  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_730
FEATURES             Location/Qualifiers
     source          1..5390
                     /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    1986..2120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2010..2029)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2064..2083
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2231..2247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2231..2247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2244..2266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2255..2271
                     /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(2279..2285,2286..2303,2304..2309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2324..2345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2462..2479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2633..3221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2713..2732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3392..4183,4184..4252))
                     /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     4015..4034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4253..4357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4425..4443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4481..4503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4603..4622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4816..4838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4830..4847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4831..4847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4874..5177
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5178..5381
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5332..5352
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5378..5390,1..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 ctgatatgaa tattggattt
     1081 atcggcaccg gcataatggg taagccgatg gcttataatt tacaacaagc cggacatact
     1141 ctttattttt cagcgcattt tgaacctgca ccgcaagagc ttattggtga acgtggcatt
     1201 gtttgctcta caccgcgtga agtcgcgcag gagtgtgagg tcatcatcac catgctccct
     1261 gataccccac atgttgagga tgttcttttt catcccaact atggcgtaat acatggttta
     1321 agccacggga aaattgtgat tgatatgagt tcaatttccc cggttgccac caaagcattt
     1381 gcacaacgta ttatcgcggg aggcgcggaa tatatcgatg cgcccgtctc tggcggtgaa
     1441 gttggcgcga aagccgggac gctttccatt atggttggcg gatgtgaaga ggtttatttg
     1501 cagatcaaac cgattctgga attgatgggt aaaaatatca ctctggtcgg taacgtcggt
     1561 gatggtcaaa cgtgcaaagt cgctaaccaa attattgttg cactgaatat tgaagcggtg
     1621 gctgaagccc tgctctttgc cagtaaatca ggtgccgatc ccgcccgcgt acgtgaagcg
     1681 ctgatgggag ggtttgcctc ctctcgcgtg ctggaagttc atggcgaacg gatgattaaa
     1741 ggaacattcg aacccggttt tcgtattagt ctgcatcaaa aagatctgaa tctggcgctg
     1801 gagaatgccc gcctgttgaa taccccgctt cctaacacgg cgacgacaca gcagttattt
     1861 agcgcctgcg ccgcgctcgg tggtaaagag tgggatcatt ctgcgttaat tcgggcgctt
     1921 gaaacacaag ccaatttcac tatccgtaaa tgaatcggta ccctcgagaa gcttgcggcc
     1981 gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2041 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2101 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2161 gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg
     2221 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2281 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2341 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2401 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2461 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2521 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2581 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2641 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2701 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2761 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2821 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2881 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2941 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3001 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3061 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3121 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4321 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4381 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4441 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4501 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4561 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4621 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4681 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4741 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4801 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
     4861 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4921 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4981 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5041 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5101 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5161 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5221 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5281 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5341 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
//
