LOCUS       sequence-209596-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_668 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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_668
FEATURES             Location/Qualifiers
     source          1..6506
                     /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    3662..3796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3686..3705)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3740..3759
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3907..3923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3907..3923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3920..3942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3931..3947
                     /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(3955..3961,3962..3979,3980..3985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4000..4021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4138..4155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4309..4897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4389..4408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5068..5859,5860..5928))
                     /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     5691..5710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5929..6033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6101..6119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6157..6179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6279..6298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6492..6506,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6506,1..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 ctgatatgaa gctctttctc tgcgaaaaac ccagtcaggg acgcgatatt gccgctgtgc
     1681 tgggtgcgac aaagaaaatg ccggggcatc aaaccggaaa cggcgtgtgc gtcacctggg
     1741 gttttggtca tctgctggag caggccagcc cggacgccta cggtgaacag ttcggcgttc
     1801 cctggcgtac tgacgtgttg ccggtattgc cgacaatgtg gcaaatgcgg gttaaaccgg
     1861 atgcagcagc tcagttctcc gtcatccaga aactgctgaa acaggctgat acggtagtga
     1921 ttgccaccga tgcggatcgg gaaggagagg tgattgctcg tgaaatcctc gattattgtc
     1981 actatcaggg aaaagttgag cggttatggc tctcggcgct ggatgaggcc agcgttcgcg
     2041 ccgcgctggc gacgataaag cccggaagta cgactttacc ctactattta gccgggcttg
     2101 gccgcgcccg ggctgactgg caaataggta tgaacatgac gcggctatat tcggtcaagg
     2161 cccgggagaa cggctacggt agcgttttat ccgtggggcg ggtgcagacg ccgacgctta
     2221 acctgatcgt ggtacgcgat cgcgatattg tgcagtttgt tccgaagccc tactggaatg
     2281 ttgtggcgca gctggcggcg ggaggaattg cgtttcaggc gcagtgggtg ccagcagagg
     2341 cgtattgtga tgaggaaaaa cgctgtatca acccgacagc ggcgcagcag gtggttcagc
     2401 tttgccagaa aaccggacag gcgacggtgg cagaggttga gaccaaaagg gaaacggagt
     2461 ctgctccgct ggcatttgat ctcggtacgc tacaacaagc ctgttcccgc cgtttcggtt
     2521 atggcgctca gcaggtgctg gatattgcgc aggctttgta tgaaacctac aaagcaacca
     2581 cgtaccctcg taccgattgc ggctacctgc caacatccat gcgtggcgag atcggacagg
     2641 tgatgtcttc gatacagcaa agcgatcccg ctttatctgc gatcatctcg cggcttgata
     2701 tgaagcatgt ttcccgtatc tggaatgaca aaaaaatcac cgcgcaccac ggtattatcc
     2761 cgacgaagca accctgtgat ctggcgaaaa tgagtgagca ggagcgcaac gtttaccagc
     2821 ttatccgtct gcattatctg gcccagttcc tgccgaacca tgaagtggat gccacgcggg
     2881 tttcgctgca ttgtggcggt cagttattta agaccagcgg taaagctgtg gtcgttgctg
     2941 gctggaagag tttgtttggt aaggatgctg acgaggatgc aggcgatgcc ggtgataaag
     3001 cacggttacc tgcaatggct aaagggagct tgtgccaggt caccggcgcg gaagcaaaac
     3061 cgcagcagac aaaaccacct gagcactata cggaagggac gttgattgca gcgatgaaga
     3121 atgcagcgca gttcgttacc gatccgcgtc ttaaaaagat cctcaaagaa aatgcggggc
     3181 tgggaacgga agcaacccgg gctggagtaa ttgaaacgct cctgaagcgt ggatttgtgg
     3241 tgaagaaagg caagcatctg ctggcgacac ctatcgcggc tgatttgatg gctgtgttac
     3301 cggcacagct taaagatccg ggtatgacgg cgctctggga gcaatcactg gatgacattg
     3361 ctgaaggccg gatgtcgctt gatgacttta tggcgaaaca gtcagcctgg acaatgcaac
     3421 tggtggcaaa ggggcagcat cagacggtta agatcacgtt accaccttct ccgccgtgcc
     3481 cgatctgcgg tggggtaacg cggcagcgca cggggaaatc aggctcattc tggggatgcg
     3541 tcaactaccc tgactgcaac gggattgtga atgtcggggc gaagaagcct ggtcgtcggc
     3601 ggaaaagtcc ggcaaagaaa agtgtctgaa tcggtaccct cgagaagctt gcggccgcgg
     3661 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3721 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3781 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3841 ggtgtgggag gttttttcgg atcctctaga gtcgatctgc aggcatgcta gcttggcgta
     3901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6061 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6121 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6181 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6241 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6301 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6361 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6421 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6481 gccagggttt tcccagtcac gacgtt
//
