LOCUS       sequence-210144-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_318 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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_318
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..106
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     98..115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     99..115
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        142..445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        446..649
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     600..620
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     646..670
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          787..883
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(944..946,947..949,950..1660)
                     /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(989..1010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1250..1269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1597..1618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3490..3624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3514..3533)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3568..3587
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3735..3751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3735..3751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3748..3770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3759..3775
                     /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(3783..3789,3790..3807,3808..3813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3828..3849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3966..3983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4137..4725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4217..4236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4896..5687,5688..5756))
                     /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     5519..5538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5757..5861)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5929..5947
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5985..6007)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6107..6126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
       61 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt
      121 cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc ctggctgacc
      181 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      361 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      421 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      481 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      541 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      601 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      661 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      721 ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa accagaaagt
      781 taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg tggtggtgca
      841 aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta cggaagtgtt
      901 acttctgctc taaaagctgc ggaattgtac ccgcgggccc accatggtga gcaagggcga
      961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1021 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1081 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac
     1141 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1201 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1321 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1381 caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt
     1441 caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1501 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1561 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1621 cgccgccggg atcactctcg gcatggacga gctgtacaag cgaattggtg gagggggcgg
     1681 ttccggtggc ggaggttccc gaattcgtcg acggagatct gatatgtctg agatccattc
     1741 gcccgccgaa gctgctagcc agcgagcttt ggaatctatg tatgcttgcc tcagtgaagg
     1801 acgaagcttc cggctggaag ctggcgctgg tgcgggtaaa acgtattccc ttattaaggc
     1861 acttcgattc ttgattgagc gaaataagga tttatttgaa aaacgtagcc agcaaattgc
     1921 ctgcatcacc tttactaacg tggcgaagga tgaaatcgtc gcgcgaacag atcgaagccc
     1981 catcgttttt tgtgatacca atcatgcgtt ctgttggtcg ttaataagca gctttcaaaa
     2041 accgttgaga gaactgattg aaacaatgcc aacatgggca gaaaagctcg aagaagttgg
     2101 tggaagcctt gggaatcgta gtattgagta taatcttgga catcgctcta ttcgcgacca
     2161 tcgtataact cttcatcatg acgatgttct ttctttaact atctcactca tggagcacga
     2221 aaaattccgt agagtgattg cagaacgctt tccaatcatt ctggtcgatg agtaccagga
     2281 cactgataaa aactggattg aggcaattaa acggcatttt cttggacaag ctagagcacc
     2341 tttatttggt tttttcggcg atcattggca aaaaatctat ggtggtggat gtggaaagct
     2401 ggaacatcag gacgtggttg aaataggtaa agaagccaat tttcgttcgg acaagataat
     2461 tgttgatgcc cttaatcgta tgagacctga gctaccgcag tgtgttaaag atcccgaagc
     2521 tacaggttta gtacgcgttt tccatacaaa caattggcaa ggagttcgtc aaaaaggcgt
     2581 gcactggagt ggtgatttac cattatgtga aggccaggct actctggaac gagttaaaac
     2641 aacgttggag caggatggtt gggtcttttc tcccaatcac accaaaattc tcatgttgac
     2701 ccatcgactt ttagccaatc agcagggcta ttcaaatata gtggcagcgt ttaaatacaa
     2761 tgaatccttc acgaaaaagg aacatgaaca tatagccttt tttgtagaca aactggaacc
     2821 cgcctgtgat gcctttaccg ctcgtaagta tggagagatg tttaatgcgc ttagcggtat
     2881 tacaccactg ttgcgtagac attcagataa ggtgagttgg cacaatgtaa tgacgcaact
     2941 tcttgctctc cgcgaaaccg gtacaattgg cgacgtaatt gagttcatcc gtgctcaacg
     3001 gaagccaaga cttccagata aggttgaaaa atttgagcag gagttactcc attttgacag
     3061 aacggctgat gttgaaatgc cccgaagatt aaaagaactt gaaaagctga aaaatattcc
     3121 ctattcggag gtcagatcac ttcgccgtta tcttgatgga tattctccat tcgaaactaa
     3181 acatggtgtg aaaggtgctg aatttgagaa tgtcctggtt gttgttgggc gtgggtggaa
     3241 ccagtataac ttcggagaaa tgttggaatt cgctggagcc caaacgattc cagtcgataa
     3301 gcaagactca tttgaacgaa accgaaattt gttttacgtc gcttgttcca gaccccaaaa
     3361 gcgccttgcg atccttttca accaagcttt gtcacctgcg gcactgacaa cacttggaaa
     3421 ttggtttgga gcagagacta taataccagc accgtgaatc ggtaccctcg agaagcttgc
     3481 ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3541 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3601 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3661 cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag gcatgctagc
     3721 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3781 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3841 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5341 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5521 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5581 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5641 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5701 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5941 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6001 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6061 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6121 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6181 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatc
//
