LOCUS       sequence-210159-        5168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_354 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5168)
  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 5168)
  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_354
FEATURES             Location/Qualifiers
     source          1..5168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..69
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     61..78
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     62..78
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        105..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          750..846
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(907..909,910..912,913..1623)
                     /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(952..973)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1213..1232)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1560..1581
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2385..2519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2409..2428)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2463..2482
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2630..2646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2630..2646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2643..2665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2654..2670
                     /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(2678..2684,2685..2702,2703..2708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2723..2744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2861..2878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3032..3620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3112..3131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3791..4582,4583..4651))
                     /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     4414..4433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4652..4756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4824..4842
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4880..4902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5002..5021
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
       61 tgtaaaacga cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgga ctctagagga tccggtacta
      721 gaggaactga aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag
      781 gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac
      841 ttctaggcct gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggg
      901 cccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      961 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1021 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1081 cccaccctcg tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac
     1141 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1201 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1261 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1321 gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag
     1381 aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag
     1441 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1501 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1561 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1621 aagcgaattg gtggaggggg cggttccggt ggcggaggtt cccgaattcg tcgacggaga
     1681 tctgatatgg tgctgacgct tgctctatat attgcatcta aagctgcggt tataacatta
     1741 cttatttatg taatgtggta ttttattttt ggtgtaaaaa atataagtga tatattaaaa
     1801 aggatcgtgc tatcaatact ttttgtctct attggtgtgc tgattctata tttgtttgga
     1861 aaagacgcat tagacctaca aataatagac aaggtcgtat atagattttc tgggttggtt
     1921 gattttatta atggaaatgg cggagacagt agttcattag agcgattttc aatttatttg
     1981 gccgtcgcaa gttttatata tagtaatatt agtatgctat tcgtaggttt tggcaatttt
     2041 agctattatg aaggaataat attgaatcac tacccgttaa gaatagcgga ctttcataac
     2101 atgcattttg aaattatcac attgtttggt cttccatttt atatattgtt aggttgtgtg
     2161 tttttttata tgtataaaat gaataaaaac atttcattta ataatactaa ggtatttaag
     2221 ttgatgatgt taactttcct tacctttctc gctatattat ctagttctgt gctcaaatat
     2281 ccatcttttt atgtttttat tatgcttttg gctgtgttgt gtgaaaataa gagtcgagta
     2341 aatgaatatt gaatcggtac cctcgagaag cttgcggccg cggggatcca gacatgataa
     2401 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2461 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2521 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2581 cggatcctct agagtcgatc tgcaggcatg ctagcttggc gtaatcatgg tcatagctgt
     2641 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2701 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2761 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2821 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2881 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2941 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3001 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3061 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3121 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3181 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3241 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3301 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3361 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3421 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3481 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3541 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3601 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3661 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3721 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3781 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3841 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3901 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3961 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4021 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4081 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4141 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4201 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4261 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4321 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4381 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4441 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4501 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4561 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4621 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4681 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4741 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4801 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4981 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5041 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     5101 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5161 tggcgaaa
//
