LOCUS       sequence-210106-        5273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_189 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5273)
  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 5273)
  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_189
FEATURES             Location/Qualifiers
     source          1..5273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     153..175
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     167..184
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     168..184
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        211..514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     669..689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     715..739
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          856..952
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1013..1015,1016..1018,1019..1729)
                     /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(1058..1079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1319..1338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1666..1687
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2596..2730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2620..2639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2674..2693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2841..2857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2841..2857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2854..2876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2865..2881
                     /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(2889..2895,2896..2913,2914..2919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2934..2955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3072..3089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3243..3831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3323..3342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4002..4793,4794..4862))
                     /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     4625..4644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4863..4967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5035..5053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5091..5113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5213..5232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
       61 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      121 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      181 cagtgagttc gagcttgcat gcctgcaggt cgttacataa cttacggtaa atggcccgcc
      241 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      301 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      361 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      421 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      481 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      541 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      601 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      661 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      721 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      781 acaccgggac cgatccagcc tccggactct agaggatccg gtactagagg aactgaaaaa
      841 ccagaaagtt aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt
      901 ggtggtgcaa atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac
      961 ggaagtgtta cttctgctct aaaagctgcg gaattgtacc cgcgggccca ccatggtgag
     1021 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1081 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1141 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1201 caccttgacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1261 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1321 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1381 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1441 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
     1501 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1561 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1621 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1681 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc gaattggtgg
     1741 agggggcggt tccggtggcg gaggttcccg aattcgtcga cggagatctg atatgattga
     1801 tacaatgaaa accttaagtg ggaaagagtt taggaaatta tttaatcgtt atgagacagg
     1861 attctatcta agtgagtttg aatacttaaa ggactttgat cctttgtttc ctgaatgcgt
     1921 agggaaagaa ttaaagttgg gttataaacg aaaagttgat gtatggtgcg aagaggatgc
     1981 tgcgctatat gcagaattta tagctacaaa agaaaggctg aaaagaggcc ggttaatcct
     2041 aaaaataaac caagtgtggt cgcgtgtata tcctgcttat atttatgcaa ttgaagaaag
     2101 aataaatgtc agggcaaagg ggagagagtc atttaagtat gaaactgtgc aagaaaaaca
     2161 atgtaaactt tccataatgt tagatggtct gaaagacctt gaatgtgaaa tagctatgat
     2221 ttcaagtgag tttgaaattg aaaaacactc tatgcactat ttttatatga tttttgatat
     2281 tgaacgctat ataaatgact gcagttactt tgttttttct gctgctaaag ttgttaaagg
     2341 cgtgattcaa tgcattggtt acattgacaa aaataacaaa gcaagttgcg atgaatttta
     2401 ttcaggcagc tatgaatgct tggatgatga tcaggaaaca ctgaactatt taaatgaatc
     2461 atgcgcagag ggtatagacg ctgataaaga tctcatcagg aagaatgagt tattagcctt
     2521 atccctgaag gaattgtatc aacttctcgt ttcaaaaaaa tgaatcggta ccctcgagaa
     2581 gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2641 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2701 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2761 caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat
     2821 gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2881 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2941 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3001 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3061 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3121 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3181 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3241 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3301 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3361 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3421 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3481 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3541 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3601 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3661 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3721 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3781 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3841 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3901 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3961 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4021 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4081 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4141 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4201 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4261 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4321 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4381 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4441 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4501 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4561 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4621 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4681 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4741 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4801 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4861 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4921 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4981 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5041 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5101 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5161 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5221 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atg
//
