LOCUS       sequence-209357-        4754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_105 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4754)
  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 4754)
  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_105
FEATURES             Location/Qualifiers
     source          1..4754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     44..66
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     58..75
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     59..75
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        102..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     606..630
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          747..843
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(904..906,907..909,910..1620)
                     /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(949..970)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1210..1229)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1557..1578
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1968..2102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1992..2011)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2046..2065
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2213..2229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2213..2229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2226..2248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2237..2253
                     /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(2261..2267,2268..2285,2286..2291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2306..2327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2444..2461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2615..3203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2695..2714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3374..4165,4166..4234))
                     /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     3997..4016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4235..4339)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4407..4425
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4463..4485)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4585..4604
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
       61 aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg tcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      661 ctccatagaa gacaccggga ccgatccagc ctccggactc tagaggatcc ggtactagag
      721 gaactgaaaa accagaaagt taactggtaa gtttagtctt tttgtctttt atttcaggtc
      781 ccggatccgg tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc
      841 taggcctgta cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccgcgggccc
      901 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      961 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1021 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1081 accctcgtga ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1141 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1201 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1261 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1321 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     1381 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     1441 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1501 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1561 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1621 cgaattggtg gagggggcgg ttccggtggc ggaggttccc gaattcgtcg acggagatct
     1681 gatatggagt ttcatgaaag tgcgatttgt gattttcgcg ctaacgcaaa ttcagtaaaa
     1741 ccacagccaa ttgcagttct ttttaaaaca atgggtgcgt gggctgtttt atgcttcgcc
     1801 gctgatgaca ctgacgcaag aatggcaata ggccaagaga tggagatgga cccgacaaac
     1861 gatgaattca taatttatgg cgctccatct aattacttac ttgatacctg caacatttac
     1921 aacaaggctg cctgaatcgg taccctcgag aagcttgcgg ccgcggggat ccagacatga
     1981 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2041 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2101 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2161 tttcggatcc tctagagtcg atctgcaggc atgctagctt ggcgtaatca tggtcatagc
     2221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3181 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3241 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3301 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4261 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4321 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4381 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     4441 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4501 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4561 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4621 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     4681 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4741 agctggcgaa aggg
//
