LOCUS       sequence-210114-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_229 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  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 5444)
  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_229
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     249..271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     263..280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     264..280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        307..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     811..835
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          952..1048
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1109..1111,1112..1114,1115..1825)
                     /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(1154..1175)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1415..1434)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1762..1783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2863..2997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2941..2960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3108..3124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3108..3124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3121..3143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3132..3148
                     /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(3156..3162,3163..3180,3181..3186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3201..3222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3339..3356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3510..4098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3590..3609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4269..5060,5061..5129))
                     /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     4892..4911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5130..5234)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5302..5320
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5358..5380)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
       61 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      121 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      181 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      241 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagttcgagc ttgcatgcct
      301 gcaggtcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      841 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      901 gactctagag gatccggtac tagaggaact gaaaaaccag aaagttaact ggtaagttta
      961 gtctttttgt cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct
     1021 cctcagtgga tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa
     1081 gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1141 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1201 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1261 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg
     1321 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1381 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1441 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1501 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
     1561 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
     1621 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1681 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     1741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1801 tctcggcatg gacgagctgt acaagcgaat tggtggaggg ggcggttccg gtggcggagg
     1861 ttcccgaatt cgtcgacgga gatctgatat gaatattgaa aatgaaactg gttttcctca
     1921 ctttcagttc gaaaaagtgg gatattatgg tgagttattc acagtagtgg tagttaatca
     1981 gacatttgat ttttcatata gcggcggtct ttgtctgatt gcggatgagc agcgactgcc
     2041 actgatgacc gactcctggt tcggcgaacc ggaaagcagc agtcttaaaa cagcaaccga
     2101 tttggtttgc agaaaagtca gggcagatgt tttattaaat ggacacgcat ggcatgccac
     2161 cggggaaacc actcgatggc aggccagttt acaggtgggt aatcttcagc gccaatttga
     2221 tatatgtggc ccttgccaat ggcaatatat atctaaccgc tggcaactta gcgaacctga
     2281 attaactgat cgggtactgt tacgccatga actggcgtca gccagtgaat ttaatcctgt
     2341 aggcattgtt attccctccc tgataacgcc tggcgataaa tcgccctgtc cagctccaca
     2401 gttatctgcc tcaccaggag ctatttgccg tagctggcct tcacgattgc gctatgccag
     2461 aggttttaca gaacagtggc aaaagactgt tcagcctttt tatccagatg aattcgattt
     2521 tgcattcatg aattattctc caccagaaca acaatacgat ggttatttaa ttggcaatga
     2581 aaaaattata ttgaacggcc tgttaccttc taaaatggaa caaacttgtt tccttccaga
     2641 tattagacta ttggcgtatt tgttttaccg ctctgataaa aacgtagaac tgcgcccatt
     2701 gttggctgac acactgacca tccacacaga cgatgagcaa ctaacactac tctggcgctt
     2761 aacgctcccc gtggattcac ttccacaacg tattgttttg acgacggagc agaggggacc
     2821 tcatggctga atcggtaccc tcgagaagct tgcggccgcg gggatccaga catgataaga
     2881 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2941 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3001 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttcg
     3061 gatcctctag agtcgatctg caggcatgct agcttggcgt aatcatggtc atagctgttt
     3121 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3181 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3241 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3301 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3361 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3421 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3481 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3541 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3601 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3661 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3721 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3781 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3841 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3901 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3961 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4021 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4081 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4141 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4201 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4261 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4321 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4381 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4441 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4501 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4561 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4621 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4681 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4741 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4801 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4861 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4921 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4981 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5041 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5101 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5161 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5221 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5281 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5341 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5401 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagc
//
