LOCUS       sequence-210050-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_728 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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_728
FEATURES             Location/Qualifiers
     source          1..6044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(829..831,832..834,835..1545)
                     /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(874..895)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1135..1154)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1482..1503
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3183..3317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3207..3226)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3261..3280
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3428..3444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3428..3444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3441..3463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3452..3468
                     /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(3476..3482,3483..3500,3501..3506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3521..3542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3659..3676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3830..4418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4589..5380,5381..5449))
                     /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     5212..5231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5450..5554)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5622..5640
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5678..5700)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5800..5819
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6013..6035
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6027..6044
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6028..6044
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag
      841 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      901 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      961 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1021 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1081 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1141 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1201 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1261 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
     1321 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1381 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1441 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1501 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgaat tggtggaggg
     1561 ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga gatctgatat gcaatattta
     1621 aaaaaacgct tttctttgat tgaactgaat gtgtttatcc cagcaatatt atttattgcg
     1681 gtaatcattc tgtgtttgac gatatatcca caagatacca gccggtatat aaatgaaata
     1741 catcattttt taacctggga aatgggcggg atttttctgg tcatgacttt tctggttgta
     1801 ctttgttgtc tgtggctggc cttctcacgt tatggcgata tcttgttagg tcagtcggga
     1861 gaaaaacctg acttcagctt gttaacctgg cttggactta tttttacttc tggaacgggg
     1921 ggcagcttgt tatatctggc ctctgtagag tggatttgga tcattcagca accgcctttt
     1981 ggcgcaacag caggaagcgc tcaggctgct cgttgggcct ctgcttatgg catgttccat
     2041 tgggggccgt ccgcatgggc atggtatttg atttgcgccg ttcctattgg ttggtttatg
     2101 catgttaaga aaacaaactc attaaaggtc agtgatttat gtcgtggatg tttgggagcg
     2161 cgtgcggatg gtttttgcgg gcattgtgtg aattttttct acatgtttgg tttgctcggc
     2221 ggcgcggtaa cgtctctggc gctgggaacg ccgatgattt ctgccgtatt ttgccatgtg
     2281 ttccatctgg atcctgccgg gcagtttatc aatgtcatgg ttatttttat ctggacgcta
     2341 gtaccattat tcattctctt ttttggactt aaaaaaggtg tggcatgggc cagtaactgg
     2401 aatattcgtg ctgacatttt gatgctactg gcaatactga tttgtggacc aacggcattt
     2461 atacttaatc aatcaattga tggtctcggc ctgatgctgc aaaattttgt agcgatgagt
     2521 ttaagtaccg atgctattgg tcgtagcgga tttccacaga tgtggaccgt attttatttt
     2581 tcatggtggg tcgtgtatgc catcccattc ggtttattta tcgcccgtat ttcaaaagga
     2641 aggacgatcc ggcaattgat tgtatgtgga actctggcag gttcattggg atgcatggtt
     2701 ttttacatgg tactggctaa tttcggctta tcgttgcaga caactcatgt catcgatttt
     2761 gttcccatac ttaacgaaca agggcgaggc gttgtcgttt ctcgtttact ggagcagcta
     2821 cccgcaagtc aggtgttttt ggttgctttt ggggctatag cattaatttc atatattacc
     2881 ggacactgta ctgtgggtta tgccctcggt tttgcgacgc aaaaacgggc agatagtgag
     2941 agtgaaccgg cattctggaa cgtggcattt tggttgatta tgactggaat cgtcgccatc
     3001 acactctatc ttcttgatgc gcagagtttg cagccgctac aaacggtctc tatcctggcc
     3061 ggactaccgc tttgcggcgt agtgtttatt ttattgaaga gttttttgac acagcttgcg
     3121 gctgaagaga aaaccgcgag agatgaatga atcggtaccc tcgagaagct tgcggccgcg
     3181 gggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3241 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3301 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3361 aggtgtggga ggttttttcg gatcctctag agtcgatctg caggcatgct agcttggcgt
     3421 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3481 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3541 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4561 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4621 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4681 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4741 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4801 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4861 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4921 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4981 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5041 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5101 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5161 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5221 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5281 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5341 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5401 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5461 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5521 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5581 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5641 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     5701 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     5761 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     5821 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     5881 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5941 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     6001 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagt
//
