LOCUS       sequence-212758-        5759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_440 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5759)
  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 5759)
  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_440
FEATURES             Location/Qualifiers
     source          1..5759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     158..175
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     159..175
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        202..505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        506..709
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     660..680
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     706..730
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          847..943
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1004..1006,1007..1009,1010..1720)
                     /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(1049..1070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1310..1329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1657..1678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3073..3207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3097..3116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3151..3170
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3318..3334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3331..3353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3342..3358
                     /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(3366..3372,3373..3390,3391..3396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3411..3432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3549..3566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3720..4308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3800..3819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4479..5270,5271..5339))
                     /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     5102..5121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5340..5444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5512..5530
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5568..5590)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5690..5709
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
       61 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt
      181 cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc ctggctgacc
      241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      601 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      661 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      721 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      781 ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa accagaaagt
      841 taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg tggtggtgca
      901 aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta cggaagtgtt
      961 acttctgctc taaaagctgc ggaattgtac ccgcgggccc accatggtga gcaagggcga
     1021 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1081 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1141 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac
     1201 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1261 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1321 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1381 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1441 caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt
     1501 caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1561 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1621 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1681 cgccgccggg atcactctcg gcatggacga gctgtacaag cgaattggtg gagggggcgg
     1741 ttccggtggc ggaggttccc gaattcgtcg acggagatct gatatgacga tgggaagagg
     1801 ttattttctg gtcaggggcg ataagaccac ctgcggcggc aaaattattg aaggggcgga
     1861 cgaccatacg atcatgggaa tcccgcaggc gcgggatatg gacagaatga cctgtggcaa
     1921 gcatccgggg atgtttatta tcgtcggcgg cgtgccggaa acggatattc acggcaggct
     1981 gatggcggga tcgctggaca gccagagcag ttgcccgtgt aaggcgcggt ttattgcgtc
     2041 gatgatggat gatgtgtatg aggtggaaac agaaaaaact gagaacaagt ctgttatgcc
     2101 cttgaaccct ttaaacaatg gaacaggaaa ttcatcacct cctgcttata cctctgatta
     2161 tcctgccctt cgtaatacgc acaatctgcc agaccagcag gtcagggata tgctagcgaa
     2221 taataatcat gatgttatgt tactgacgta tgaagagggc gctgaagttc tgcaatcgtg
     2281 gggttgtcag gattttaagc ggaaatgggt tgacatcacg caatctgcac cagggcagtt
     2341 aattattaat tatggcgtaa atggtaaaga tgtagtgaca acttcaatcg tgcttgcccg
     2401 cctgggggat tttggtatca aagcaactgc ctatgttaat cataaaggaa ccgaactgat
     2461 aaaacttaca ggatatcccg gcattcggaa aatactgaat gcaccggtgt ttgcagctaa
     2521 aaaccccaaa atagtagatg ttggaattgg gaaatatgga ctcacaaact ctatagtaaa
     2581 aggggcaaga ttaacgtttt acgtcgcagc tgcttataga actattgatt tcatcctgaa
     2641 tgatgaaaca tcgttggctg agtttatagg tgcgttggca acagatgttg gtaaaattgc
     2701 gatagcttca gcagttagtt ggggcgcagg aaaggcgtta ctggccactg ggtttgttgc
     2761 tggacctttg gttggggtag tggttgtggg gatcggcgcg gcccttctat taaactatct
     2821 tgataataaa tttggtttaa ctgatcaagt tgtcgcttat attgaagcgg ctcagcaaga
     2881 atttgttgag aaggcaagag agattgaaga agggatttgg gatctgggag ccatgtgggc
     2941 aaaacacatg cttattaaag ggaaggaagt actggaacat gaagtgaagg agtatttaaa
     3001 agataaacta aataatgtga gtaaatggga attgaaatga atcggtaccc tcgagaagct
     3061 tgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     3121 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3181 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3241 gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgatctg caggcatgct
     3301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5461 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5521 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5581 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5641 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     5701 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggag
//
