LOCUS       sequence-210335-        5207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_673 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5207)
  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 5207)
  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_673
FEATURES             Location/Qualifiers
     source          1..5207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          132..228
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(289..291,292..294,295..1005)
                     /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(334..355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(595..614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     942..963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1806..1940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1830..1849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1884..1903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2051..2067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2051..2067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2064..2086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2075..2091
                     /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(2099..2105,2106..2123,2124..2129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2144..2165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2282..2299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2453..3041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2533..2552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3212..4003,4004..4072))
                     /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     3835..3854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4073..4177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4245..4263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4301..4323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4423..4442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4636..4658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4650..4667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4651..4667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4694..4997
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4998..5201
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5152..5172
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5198..5207,1..15)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
       61 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      121 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      181 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      241 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag
      301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      361 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      481 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      541 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      721 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
      781 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      841 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
      901 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      961 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgaat tggtggaggg
     1021 ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga gatctgatat gaaaaagctc
     1081 aaagttgtgt tgttgtcggg agtgttgaca gtgtcaacag tcgcactcgc tgcaccttca
     1141 gtgaatcagg aaaacagcct gacggtgaat acgtcgttac agatgctgga acagcaacaa
     1201 atgacgcgga gcgcacagga gtgggggctg acagcggaag agtggcagcg ttatgaaaca
     1261 ctgaaaaaag gacggcgcgg cgtgctgtct ccggggttag atcccctgac gatgctgggc
     1321 atcgaggcgc gtagtgacga agaacgtcgc catttcgctg aactggcggt gaagcaggag
     1381 tttcagcggg ttgaagcgga actggcattt cagcgggaag tgaacagcgc gtggacgcgg
     1441 gtctatcccg gtgttctgcc cgttcaggat ctgcggagtg aagtcagtaa cgcccgtcag
     1501 gcgttgtttg tgaaaggcag ctgtccggcc tgcgagcgca agctgactca gttgatgaaa
     1561 tcgaaccagc cgctggatat ctacctcgtt gatagcggag gtaaggatga gacggtccga
     1621 aactgggcga agaagcacaa tatcccggct gaaaaggtga aatcccgcct gatcacgctg
     1681 aaccacgaca atggtatgtg gctaaaatat ggcagtgggc tgatgccggt tgtattacaa
     1741 caggggccgc agggatggca gcgcgtaaac tgaatcggta ccctcgagaa gcttgcggcc
     1801 gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1861 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1921 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1981 gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg
     2041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3181 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3241 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3301 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3361 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3421 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3481 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3541 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3601 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3661 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3721 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3781 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3841 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3901 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3961 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4021 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4081 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4141 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4201 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4261 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4321 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4381 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4441 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4501 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4561 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4621 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
     4681 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4741 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4801 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4861 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4921 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4981 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5041 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5101 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5161 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgttta
//
