LOCUS       sequence-210125-        4796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_259 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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_259
FEATURES             Location/Qualifiers
     source          1..4796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..119
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     111..128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     112..128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        155..458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        459..662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     613..633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     659..683
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          800..896
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(957..959,960..962,963..1673)
                     /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(1002..1023)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1263..1282)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1610..1631
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2063..2197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2087..2106)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2141..2160
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2332..2348
                     /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(2356..2362,2363..2380,2381..2386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2401..2422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3469..4260,4261..4329))
                     /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     4092..4111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4330..4434)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4502..4520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4558..4580)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4680..4699
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
       61 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      121 cggccagtga gttcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
      181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      661 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      721 gaagacaccg ggaccgatcc agcctccgga ctctagagga tccggtacta gaggaactga
      781 aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc
      841 cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct
      901 gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggg cccaccatgg
      961 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1021 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1081 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1141 tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1201 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1261 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1321 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1381 tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca
     1441 tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1501 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1561 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1621 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcgaattg
     1681 gtggaggggg cggttccggt ggcggaggtt cccgaattcg tcgacggaga tctgatatgg
     1741 atataacacc tttccttcat gcgctttgtg ctgtggctgc gcagctactg attggtcttt
     1801 ttaccgggaa ctgggcttac ggggcgatag ccggttgtac gttcttcatt gcgcgtgaac
     1861 acacccaggc agaatatcgc tggattgaaa tgttcgggca tggcaagcga atgaatatgc
     1921 cgtggtgggg cggttttgat ccgcgcgcgt gggatgtggc aagcctgatg gattttgctg
     1981 tgccggtggt ggcgtgtctg ctggtctggc tgttggttaa tcgtgggtga atcggtaccc
     2041 tcgagaagct tgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca
     2101 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2161 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2221 tatgtttcag gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgatctg
     2281 caggcatgct agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4441 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4501 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4561 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4621 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4681 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4741 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgtt
//
