LOCUS       sequence-209689-        6884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_722 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  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 6884)
  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_722
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        44..347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        348..551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     502..522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     548..572
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          689..785
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(846..848,849..851,852..1562)
                     /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(891..912)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1152..1171)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1499..1520
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4040..4174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4064..4083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4118..4137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4285..4301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4285..4301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4298..4320)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4309..4325
                     /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(4333..4339,4340..4357,4358..4363))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4378..4399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4516..4533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4687..5275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4767..4786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5446..6237,6238..6306))
                     /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     6069..6088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6307..6411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6479..6497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6535..6557)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6657..6676
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6870..6884,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6884,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca taacttacgg
       61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      541 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      601 acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat ccggtactag
      661 aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg
      721 tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact
      781 tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt acccgcgggc
      841 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      901 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      961 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1021 ccaccctcgt gaccaccttg acctacggcg tgcagtgctt cgcccgctac cccgaccaca
     1081 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1141 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1201 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1261 ggcacaagct ggagtacaac tacaacagcc acaaggtcta tatcaccgcc gacaagcaga
     1321 agaacggcat caaggtgaac ttcaagaccc gccacaacat cgaggacggc agcgtgcagc
     1381 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1441 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1501 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1561 agcgaattgg tggagggggc ggttccggtg gcggaggttc ccgaattcgt cgacggagat
     1621 ctgatatggt tgccattgtt attgtttctc atagtctacg tctcgcccag gggattgagg
     1681 aattagcact acagatgtct ggaggggacg ttcctctggc cattgctgca ggtattgatg
     1741 atccgcaaaa tccgattggc actgatgcta ttgccattat gtctgcgatt gaatcggtct
     1801 ggtcaccgga tggcgttctg gtactcatgg atatgggcag tgcgctgctt agtaccgaga
     1861 tggcgctgga gctgctttct gaagagcaaa gaagtgcaat ttatctgctc gccgcccctg
     1921 tagtggaagg tgccatgtca gcggtaacgt ctgccgcagc ggggctttcg gtgacggaga
     1981 ttatcgctga agtggatctt gcgctgtgcg ctaaacagca gcagctcatg ccatccccgc
     2041 ctgcaggcga ggttattcca gcaatatcac cagtcaatca ttccggtgaa tggcaaactt
     2101 tttgctggac gatatgtaat ccccacggca ttcatgctcg tccggcagcc agcatcctga
     2161 aagtatcagc acaatattcg gcaaacatta tcgtcatcaa aggcgataaa cgcgccagca
     2221 cccgcagtct caacgaactg gcaatgctgg gcgtccgctg tggcgatgaa atcatcttcc
     2281 aggccgaagg caccgatgcc agtgacgctc tcaaggcaat tgaaacgttg gcgaaaaatc
     2341 attttggtga agctcatctg ctcgcttctt cagagattcc gctaacggaa acgccggtgg
     2401 tcagcgcccc tgtcgatggc gcaatatcag gcttgtcagt acaaaatggt atcgctattg
     2461 gtcctgttaa atggtttacc tgcgaacgac cagaaattac gcagcgcacc gtcgattctc
     2521 cgcaagaaga gctgtcgcga atcgagtctg ctatcgacat tgttgtttgt gaacttgccg
     2581 ataaagcggc aggccctgaa ggcgatattt ttgccgccca taaaatgatg cttgaagatc
     2641 cagagattaa ccggcaatta cagcaacgtc tggcaaaggg caaacaggcg gaattcgcat
     2701 ggttagaagt catgcaggca ttagcagagc aatattgtca ggcggaaaca ctgtatttac
     2761 gtgaacgcga ggcagatatc cgtgacctta cacgtcaggt actgaaccag ctttgcgggg
     2821 taagtgaaca acactttatc actaccgcac cctgcatttt gttagccaac gatctgttgc
     2881 catcacagat aaccagcctg aataaagcac acattcttgg catttgtctt cataatggcg
     2941 gaacaacttc acatacagcg atccttgcca gagcaatggg aatccccgct attgtgaaag
     3001 cggcgattac cccgcaaaac gtccgcgata acgatacggt cattcttgat ggtgaaacag
     3061 gccgtttgtg gctacagccg gatgaggtca cccgcctgga tttactgcaa cgcgccgaag
     3121 catggcggca acaaagggat cgtcagttag ctgatgccat gctgcccgct gtcacgcaag
     3181 gtgggcgtaa aatcagcgtg ctggcgaata ttggtgatct gcaagatatt gaggctgcgc
     3241 tatctcacgg ggctgagggc gtgggtttac tgcgcacaga gtttttattc catgagtctg
     3301 caacattacc cgatgaagaa gagcagtttc gcgtgtattg ttctgttgct caggcttttg
     3361 gcgataaacc cgtcaccatc cgcactctgg atattggcgg tgacaagcca ttaccttcgt
     3421 accctttacc cgcagaggac aacccgttcc tgggacttcg cggtatccgg ttgtgcctgg
     3481 cgcatccgca aatatttatt ccgcaattac gcgcacttct ccgggccggg aaagaatacc
     3541 caacgctgca aattatgctg ccgatggttt caacgctcga agaggtcaac gccgtcaaaa
     3601 cgctgataca aacgcaggca aaactgctgg gacttactgc agaaaatctt ccggcgctgg
     3661 gaattatgat tgaggttcct gccgcagtca tgattgcaga caaattagcc agtgaagtcg
     3721 atttcttttc catcggcacc aacgatctga cgcaatatat tatggcggca gacagaggaa
     3781 atagcaccgt cgcgaaactg gtcgattatt gtaatgacgc ggtgattaac gctattgcta
     3841 tggtgtgtca ggccggacgt aataatgaaa ttccggtcag tatgtgcggc gaaatggctg
     3901 gagatatcca gcaaacagca cgactgctca ccatggggat tgataagctc agtgccagtc
     3961 catcccggct tccggccttg aaagcagcaa tcagaacctc ccactgaatc ggtaccctcg
     4021 agaagcttgc ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     4081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4141 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4201 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     4261 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4321 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4381 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5401 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5461 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5521 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5581 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5641 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5701 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5761 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5821 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5881 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5941 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6001 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6061 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6121 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6181 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6241 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6301 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6361 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6421 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6481 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6541 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6601 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6661 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6721 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6781 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6841 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
