LOCUS       sequence-209746-        5549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_034 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5549)
  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 5549)
  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_034
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..97
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     89..106
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     90..106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     637..661
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          778..874
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(935..937,938..940,941..1651)
                     /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(980..1001)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1241..1260)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1588..1609
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2794..2928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2818..2837)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2872..2891
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3039..3055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3039..3055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3052..3074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3063..3079
                     /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(3087..3093,3094..3111,3112..3117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3132..3153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3270..3287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3441..4029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3521..3540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4200..4991,4992..5060))
                     /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     4823..4842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5061..5165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5233..5251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5289..5311)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5411..5430
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
       61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc
      121 atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
      661 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
      721 cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag ttaactggta
      781 agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga
      841 actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct
      901 ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg aggagctgtt
      961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt
     1141 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1381 caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg
     1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1621 gatcactctc ggcatggacg agctgtacaa gcgaattggt ggagggggcg gttccggtgg
     1681 cggaggttcc cgaattcgtc gacggagatc tgatatgata gaaatgaaaa ataacacgcc
     1741 atttcctttg ttgtcatttg aaaaatatgg tcgttatggt cttttgtttg atgttattgc
     1801 tattaaaatg tcattgcgaa ttaaaaatgg attttatgct gatcttgctg agtttcaaaa
     1861 agaattgtcc atgtctgatg aatactatgg ggagtctgaa acaagtagtc taaaatctga
     1921 aacagactta gttctttgta aaagaaatac agatattcac gtgacaggta gtgctcatgc
     1981 cccttcaggg gataagtctc agtggaaggc ttgtgtgagg gttaatagtt tttctaaaga
     2041 actgagttta tctggtgtcc gctacttaca atatgagcga aatcgttggc agatgagctt
     2101 accagataaa attattaatg ttcctctacg ctatgagctt gcttacggtg gtatttggca
     2161 accagatggt atggagaaat tggttttttc agcaaaccct gtaggttgtg gatattatcc
     2221 tgatatttct caacttaata caagttgtca atataaactc ccacaaataa ccagctctgc
     2281 tttgtcagaa aacgctacca tttttaatgg ggaatctgac ttttttcaag gtgttggccc
     2341 tgtatctcga tggtggaaaa gtcgcctcca gtatgcagga acctacaatg aagtatggcg
     2401 tgaaaagcgc tatccctatt tgccggatga ttttgatgag cggttctaca atagtgctca
     2461 tccggatatg atatacacgg ggtttctctc tggcgatgaa aacatctctc ttgaaggttt
     2521 ttttaaattt gaacaggttg ttaagacaaa attgccaggt atacgacctg tacttatatt
     2581 aaaaaccaag aataatacat cacatatgtt cttacctgtt gcggatacaa tggttatcga
     2641 tttaagtcga caggaaatat accttacctg gagattaaca atacctgatt tttttggaat
     2701 gaaagaagga gtgctgagtt gtattattcc tgaatgtata gggaaaaaat attatgggtg
     2761 aatcggtacc ctcgagaagc ttgcggccgc ggggatccag acatgataag atacattgat
     2821 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2881 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2941 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta
     3001 gagtcgatct gcaggcatgc tagcttggcg taatcatggt catagctgtt tcctgtgtga
     3061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5161 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5221 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     5281 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5341 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5401 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5461 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     5521 aactgttggg aagggcgatc ggtgcgggc
//
