LOCUS       sequence-210328-        5255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_666 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5255)
  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 5255)
  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_666
FEATURES             Location/Qualifiers
     source          1..5255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          131..227
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(288..290,291..293,294..1004)
                     /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(333..354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(594..613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     941..962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1853..1987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1877..1896)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1931..1950
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2098..2114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2111..2133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2122..2138
                     /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(2146..2152,2153..2170,2171..2176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2191..2212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2329..2346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2500..3088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2580..2599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3259..4050,4051..4119))
                     /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     3882..3901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4120..4224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4292..4310
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4348..4370)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4470..4489
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4683..4705
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4697..4714
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4698..4714
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4741..5044
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5045..5248
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5199..5219
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5245..5255,1..14)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
       61 gggaccgatc cagcctccgg actctagagg atccggtact agaggaactg aaaaaccaga
      121 aagttaactg gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg
      181 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag
      241 tgttacttct gctctaaaag ctgcggaatt gtacccgcgg gcccaccatg gtgagcaagg
      301 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      361 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      421 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      481 tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      541 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      601 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      661 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      721 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
      781 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      841 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      901 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      961 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcgaatt ggtggagggg
     1021 gcggttccgg tggcggaggt tcccgaattc gtcgacggag atctgatatg tcagacaaga
     1081 cagaaaagaa agctggtgcg ctgcattctg agctgcgcat cgagctgcat acccaatatg
     1141 ccattaacac ctggagcggg cggccagcct cggatagcaa gccttcgctc atcggaatgc
     1201 cgctgttttt ccggctcatc tcccggatta accgcgattc actggcagat aatccgtggg
     1261 cggatgcggc cctggtggaa gtggaaacat tactggagac ggccagccag cagctccaga
     1321 aatggttgaa tgatatcgat gccctgatgg cgatgctgcc agctaacgct tccatttccg
     1381 gcgtcacctc cacctcgccg ctgaatatcg gcgttcacag ccataccccg ctgggatacc
     1441 gctgcgttta tcttcttgtt gggtttgatc agcttattct gcgtgtcttc caggctcacc
     1501 attacggtct gatgtcgttg aaagagcgca aagcctggtt acatcgtggt agccatcaaa
     1561 ttcgccgcat tttctcggtt gccctgcgtt accgttctca ccacgttacc cgccaggaca
     1621 ttgcctcaaa taacgatgct gcccgacagg ctatcgaaca atttggggaa ctcgacaggg
     1681 cgatcctgct gggaacccgc cgttcccgat tttctccgcc agtaagcgcc gaaagtatta
     1741 aggcgctgaa agccgccttt aaacgtctgt cacgcaaaga gaaagcggca aagcaggagg
     1801 tgcgggatga tccgcaatga atcggtaccc tcgagaagct tgcggccgcg gggatccaga
     1861 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1921 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1981 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2041 ggttttttcg gatcctctag agtcgatctg caggcatgct agcttggcgt aatcatggtc
     2101 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2161 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2221 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2281 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2341 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2401 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3061 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3361 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4261 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4321 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4381 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4441 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4501 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     4561 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4621 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     4681 ttcccagtca cgacgttgta aaacgacggc cagtgagttc gagcttgcat gcctgcaggt
     4741 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     4801 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     4861 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     4921 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     4981 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     5041 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     5101 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     5161 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     5221 acggtgggag gtctatataa gcagagctcg tttag
//
