LOCUS       sequence-210220-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_469 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  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 5669)
  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_469
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..92
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     84..101
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     85..101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        128..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     632..656
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          773..869
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(930..932,933..935,936..1646)
                     /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(975..996)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1236..1255)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1583..1604
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2909..3043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2933..2952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2987..3006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3154..3170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3154..3170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3167..3189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3178..3194
                     /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(3202..3208,3209..3226,3227..3232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3247..3268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3385..3402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3556..4144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3636..3655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4315..5106,5107..5175))
                     /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     4938..4957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5176..5280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5348..5366
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5404..5426)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5526..5545
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
       61 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc
      121 tgcaggtcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      661 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      721 ggactctaga ggatccggta ctagaggaac tgaaaaacca gaaagttaac tggtaagttt
      781 agtctttttg tcttttattt caggtcccgg atccggtggt ggtgcaaatc aaagaactgc
      841 tcctcagtgg atgttgcctt tacttctagg cctgtacgga agtgttactt ctgctctaaa
      901 agctgcggaa ttgtacccgc gggcccacca tggtgagcaa gggcgaggag ctgttcaccg
      961 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1021 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1081 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt
     1141 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1201 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1261 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1321 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaagg
     1381 tctatatcac cgccgacaag cagaagaacg gcatcaaggt gaacttcaag acccgccaca
     1441 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1501 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
     1561 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1621 ctctcggcat ggacgagctg tacaagcgaa ttggtggagg gggcggttcc ggtggcggag
     1681 gttcccgaat tcgtcgacgg agatctgata tgcatgtcat gacttcagat aataaaaata
     1741 ttgatttaca attaaatata aatgagcatg gagtttctca taaactgatt aaaaaaacgt
     1801 ctaataaatt gctcgtcttt ttttcaggta cagataaaaa agatggccgc tttgattttt
     1861 ggaaatcagc agatgaactg ccatataaca ttttgttaat taacaatggt aaaaatgaat
     1921 ggtaccaaga tagtattcca gggtttagtt cttcaatatc agaaacaata gaaaaaataa
     1981 aatatatttc tgaaaaacta ggttgtgacg agattataac aataggcgtc agtatgggag
     2041 gatatgcagc ttcattattt ggcgctcttc tggattgtcg tgtgttagcc tttagctttg
     2101 acactgtact aaaatatccc ttgagccgta gtgctaaaag aattcctaaa aagacaaaaa
     2161 ttatatataa gaatctacgc cctattataa aaaattctaa atgtagaatt acagcattaa
     2221 gtggggaaat ggattttcct gatttacttt cattgagtag gatttcagat cttaaaaatg
     2281 ttaaagcgta ttccgttcga ggagtaacac atggtgtagg gcggtttata gataaaagat
     2341 atggaattcc taatataata accttttttg tggaaaataa tacacttccc gaaattaaag
     2401 agattaataa tctttgccat aataaaatct tatcaaaaaa tatattcctg tcttaccaag
     2461 catttgttaa caaagatttt tccactgctc aatctttaat cagagaggct ttgttggctg
     2521 aaccattact tgaacctgct atatttattt ctgctttagt aaatatggaa ctcaagaact
     2581 atactttagc tgtagagcag tttgctttta ttgtcggtat ttcgccacat tttacaacag
     2641 ctaaatataa tttggcaaag tcattaagga tgtcaaaaaa atatgatcag gcaatacaac
     2701 atttctatga atatatatct ttaatgcctt catccgctgg ctcatattat aatttatctc
     2761 taatatatga aagaaaaggg ttattggaag atgcaaaaaa aatggcccaa atggcatata
     2821 gtctatcccc tgaaagtgaa acatataaaa aaaggtatga tacttatttt tcatgaatcg
     2881 gtaccctcga gaagcttgcg gccgcgggga tccagacatg ataagataca ttgatgagtt
     2941 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3001 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3061 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttcggatc ctctagagtc
     3121 gatctgcagg catgctagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3181 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3241 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3301 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3361 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3421 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4081 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4141 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4201 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4261 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4321 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4381 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4441 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4501 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4561 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4621 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4681 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4741 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4801 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4861 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4921 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4981 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5041 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5281 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5341 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5401 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5461 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5521 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5581 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     5641 ttgggaaggg cgatcggtgc gggcctctt
//
