LOCUS       sequence-210272-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_658 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  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 5378)
  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_658
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          112..208
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(269..271,272..274,275..985)
                     /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(314..335)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(575..594)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     922..943
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1957..2091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1981..2000)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2035..2054
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2202..2218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2202..2218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2215..2237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2226..2242
                     /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(2250..2256,2257..2274,2275..2280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2295..2316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2433..2450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2604..3192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2684..2703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3363..4154,4155..4223))
                     /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     3986..4005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4224..4328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4396..4414
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4452..4474)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4574..4593
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4787..4809
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4801..4818
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4802..4818
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4845..5148
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5149..5352
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5303..5323
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5349..5373
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
       61 gactctagag gatccggtac tagaggaact gaaaaaccag aaagttaact ggtaagttta
      121 gtctttttgt cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct
      181 cctcagtgga tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa
      241 gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      301 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      361 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      421 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg
      481 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      541 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      601 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      661 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
      721 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
      781 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
      841 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
      901 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
      961 tctcggcatg gacgagctgt acaagcgaat tggtggaggg ggcggttccg gtggcggagg
     1021 ttcccgaatt cgtcgacgga gatctgatat gcatgtaatt tcagtaattt ctactaaagg
     1081 tggtgaaggt aaatccactc attccgccaa cctggctggc ttctgtgccg atgccggtct
     1141 caaaaccctc ctgattgacg gcgactacgc ccagcccacg gtttccagtt actatccttt
     1201 gcgctacgaa gcgccatgtg gtttgtacga gttacttatg cagactgtgg atcttaatcg
     1261 gccggaacaa ctgatttccc acactgccat tgataacctt gatctgatta tttctaacga
     1321 tccttatggt cagttaaaaa tggcaatgat gcacgctccg gacgggcgca tcagactacg
     1381 taaccagctt cagcatccat tgtttgcctc ttatgacgtg attatcagtg atacgcaagg
     1441 cgcgcgctca atcatgttag agctgatgtt gttggcgaca acacatacgg cactggcggt
     1501 tgtgaaaccg gtaataccgg acgttcgcga gtttctacgg ggaacggtaa cgctgatgga
     1561 ggatctactg ccttaccgtg gatttggtat taccttgccg tcaatgaaaa tcctggtcaa
     1621 ctgcatggat tacaccctgc tggctgaaga agcactgggt gcgttaacta cgattattgc
     1681 tgatgggacg tatagcaaag cggatattcc ccccgtcact ctgctcgaaa cccaaatcta
     1741 cgatcttgaa gtctacaagt taggccatac gcgggggcaa cccgctcacc gtcttgaata
     1801 ccataccgat cgtaaaagtt tgccagccgc tcagactatg cacgatctgg cctgcgaatt
     1861 atttcccgaa tggcgcgacc gttttgatgc cgtactgcaa aaccagccga agggggataa
     1921 atgaatcggt accctcgaga agcttgcggc cgcggggatc cagacatgat aagatacatt
     1981 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2041 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2101 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct
     2161 ctagagtcga tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2221 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2281 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2341 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2401 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3301 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3361 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3421 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3481 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3541 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3601 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3661 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3721 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3781 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3841 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3901 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3961 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4261 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4321 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4381 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     4441 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     4501 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     4561 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     4621 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     4681 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     4741 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     4801 tgtaaaacga cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg
     4861 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4921 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4981 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     5041 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     5101 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     5161 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     5221 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     5281 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5341 ataagcagag ctcgtttagt gaaccgtcag atcgcctg
//
