LOCUS       sequence-210482-        5585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_725 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  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 5585)
  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_725
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          136..232
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(293..295,296..298,299..1009)
                     /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(338..359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(599..618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     946..967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2188..2322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2212..2231)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2266..2285
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2433..2449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2433..2449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2446..2468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2457..2473
                     /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(2481..2487,2488..2505,2506..2511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2526..2547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2664..2681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2835..3423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2915..2934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3594..4385,4386..4454))
                     /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     4217..4236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4455..4559)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4627..4645
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4683..4705)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4805..4824
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5018..5040
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5032..5049
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5033..5049
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5076..5379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5380..5583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5534..5554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5580..5585,1..19)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
       61 acaccgggac cgatccagcc tccggactct agaggatccg gtactagagg aactgaaaaa
      121 ccagaaagtt aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt
      181 ggtggtgcaa atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac
      241 ggaagtgtta cttctgctct aaaagctgcg gaattgtacc cgcgggccca ccatggtgag
      301 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      361 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      421 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      481 caccttgacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      541 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      601 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      661 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      721 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
      781 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      841 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      901 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      961 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc gaattggtgg
     1021 agggggcggt tccggtggcg gaggttcccg aattcgtcga cggagatctg atatggataa
     1081 gattattata agtccggcta aatatatcca gggaaatggt tctctggata atattgctac
     1141 atatgcggcg attttgggaa cagaaccatt aattattgca gatgaatttg tgaccggtct
     1201 ggtgggtgac agagtaagcc agagctttgc aaaagaaaat attactgcag attttgatat
     1261 cttttgtggt gaatgctcgc aaaatgaaat ttctcgaatt cgtgagaaat ttaatcagcg
     1321 taaatataat gtcgttattg gtattggcgg cggtaaaacg ttggatacgg caaaagccgt
     1381 agcgtattac caaaaaattc cggtggtggt ggtgccgaca attgcttcca cagatgcgcc
     1441 aaccagttct ctggcagtaa tttacacccc ggaaggtcag tttagcgaat atctgttttt
     1501 cccgaaaaac ccggatatgg tgattatgga tactggcgtc atttcagccg cgccagtgcg
     1561 tctgctggtg gccggtatgg gagatgcatt gtcaacctgg tttgaggcga gagccaatca
     1621 ggcgtcagga aaagccacaa tggcaggtgg ggcgtcgaca ctggcggcac tggctattgc
     1681 ccgactctgt tataccacgc tactggaaga tggctataag gcaaaagtcg cggttgaaca
     1741 aggtgttagc accaaagcgg tagagaatat tatcgaagct aatacctatc ttagcggcat
     1801 tggatttgaa agtagcggcc tggcggcagc tcatgccatt cataacggca tgacgcaact
     1861 ggaggaatgc catcattgct accacggtga aaaagtggca tttggcgtcc tggtacagct
     1921 cgtactggaa aacgcaccac aggaggaaat tgaaaccgta ctcaatttct gccatagcgt
     1981 tggtctgccg acaaatcttc atatgttagg cgtgaaggaa attaataaag ataaattacg
     2041 ggcagtggct gaagccgcga ctgctgaagg ggaaactatc cataatatgc cattcgtggt
     2101 cactgcgcag gacgttcttt gcgcaatttt aactgcacat catttgggac tttgaatcgg
     2161 taccctcgag aagcttgcgg ccgcggggat ccagacatga taagatacat tgatgagttt
     2221 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2281 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2341 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttcggatcc tctagagtcg
     2401 atctgcaggc atgctagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3241 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4561 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4621 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4681 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4741 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4801 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4861 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4921 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4981 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5041 acggccagtg agttcgagct tgcatgcctg caggtcgtta cataacttac ggtaaatggc
     5101 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5161 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5221 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5281 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5341 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5401 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     5461 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     5521 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     5581 gctcg
//
