LOCUS       sequence-209341-        5159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_064 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5159)
  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 5159)
  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_064
FEATURES             Location/Qualifiers
     source          1..5159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        24..327
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        328..531
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     482..502
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     528..552
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          669..765
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(826..828,829..831,832..1542)
                     /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(871..892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1132..1151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1479..1500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2295..2429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2319..2338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2373..2392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2540..2556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2540..2556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2553..2575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2564..2580
                     /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(2588..2594,2595..2612,2613..2618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2633..2654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2771..2788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2942..3530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3022..3041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3701..4492,4493..4561))
                     /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     4324..4343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4562..4666)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4734..4752
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4790..4812)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4912..4931
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5125..5147
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5139..5156
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5140..5156
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgagcttg catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga
       61 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      121 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      181 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      241 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      301 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      361 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      421 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      481 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      541 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      601 gaccgatcca gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa
      661 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
      721 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
      781 ttacttctgc tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc
      841 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      901 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      961 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg
     1021 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1081 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1141 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1201 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1261 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
     1321 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1441 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1501 accgccgccg ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc
     1561 ggttccggtg gcggaggttc ccgaattcgt cgacggagat ctgatatgct gaaagcacag
     1621 ctcgatgaat ttgttgttgg catgcattct gccgacccgg ttatcaggat cgccagatta
     1681 atatcgcttg aagagattaa ccgccatcag gattttttcg aacactgcgc gaaagaatat
     1741 cgtaagctgg cgacagagct tatatttgca ttggctgaac aactgaacgt tgagatggcg
     1801 gaaaataacc cgctagttac ctttgccccc ttcaaatgca accgtaagcg aaaaggcaaa
     1861 atgggaaagt ggcagtattg ttttcacggt tttcattgta catttgagaa taaaaaaaca
     1921 gagcaaaata ttgaagttcc cctcgcctac ggctttgatt tcggcgatct ggacccgtat
     1981 tttttttccg gatttatcaa atcgaccccc gcctggcagc ctctgccagt ggctatttac
     2041 gatgattacc acgacggtag caggatcatt caacaactgc tggcgctggg aaagttacaa
     2101 aaaatccctt caccgatccc acaatacacg ggggttgctg cggtagatcg gtccaatgtc
     2161 gacatcgcaa attttcgctc aacactggag agtcgcctgc accgatgtaa attacgctgg
     2221 cttttgaagc atgtaaaaaa ttcacaaagg agcgaaagat gaatcggtac cctcgagaag
     2281 cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     2341 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2401 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2461 aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg
     2521 ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4681 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4741 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4801 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4861 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4921 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4981 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     5041 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     5101 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgag
//
