LOCUS       sequence-210453-        6032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_695 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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_695
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          133..229
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(290..292,293..295,296..1006)
                     /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(335..356)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(596..615)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     943..964
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2632..2766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2656..2675)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2710..2729
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2877..2893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2877..2893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2890..2912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2901..2917
                     /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(2925..2931,2932..2949,2950..2955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2970..2991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3108..3125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3279..3867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3359..3378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4038..4829,4830..4898))
                     /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     4661..4680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4899..5003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5071..5089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5127..5149)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5249..5268
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5462..5484
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5476..5493
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5477..5493
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5520..5823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5824..6027
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5978..5998
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(6024..6032,1..16)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
       61 ccgggaccga tccagcctcc ggactctaga ggatccggta ctagaggaac tgaaaaacca
      121 gaaagttaac tggtaagttt agtctttttg tcttttattt caggtcccgg atccggtggt
      181 ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt tacttctagg cctgtacgga
      241 agtgttactt ctgctctaaa agctgcggaa ttgtacccgc gggcccacca tggtgagcaa
      301 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      361 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      421 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      481 cttgacctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
      541 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      601 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      661 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      721 caactacaac agccacaagg tctatatcac cgccgacaag cagaagaacg gcatcaaggt
      781 gaacttcaag acccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
      841 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
      901 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
      961 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagcgaa ttggtggagg
     1021 gggcggttcc ggtggcggag gttcccgaat tcgtcgacgg agatctgata tgacggctga
     1081 cagttatctg gaaatggtgc tggttctgat gggctggctg atcaacaacg cgctgtggac
     1141 gattattacc accacgggca ttttcgctat tccgctggcg ttcaaaatcg ccggtgtctg
     1201 gctgaaggtt cgggaagagg gagatgatga agggaataaa ggggcgctgg cattaccgcg
     1261 tatagagcat gtcatctatg tcgcttttgt ggtgatcatg ttctgtgcga tgccaataca
     1321 ggatgttgat atctcgacga tgaagtttga tcgcacccgt tcttcccagt gtggtgtgaa
     1381 tgtacctaag ccagatgagt ctggctatgg gccactgatt gccgagctta acggccagac
     1441 tgcgaaaatc ccggcatggt ggtatctggt acattcgcta tcgaaagggg tgacgcacgc
     1501 cgcgatagcc tctattccct gtggcaatga cttccggatg ctgcgttttg aagtgcaaca
     1561 tacgaagctg attgatccga ttattcgcca ggaggttcag gagtttgcca gccagtgtta
     1621 tgcgaagtcg tattacaagc tgaagaacgt gaattcaggg ctgtcagcgg agacggtaga
     1681 ttccgttggc tggattggtt cggatttttt tctcaatact ggcggctatt acgattacta
     1741 cacgtcaacg atgccccggg cggcgtggcc ttacagcgac agccgggaca gtggttatcc
     1801 aaatgttggc cgtggtggtt atccaacctg taaagactgg tggagtgcat cagggaaagg
     1861 gctacaaagc cgtctgctga aaaacttcga tacccgaacg atcaatatga tgaaggcaaa
     1921 atcccccaat aactgggaag aaagcctgtt gcgctggctg gtcagtcctc agaatacaca
     1981 actttctggt ggtggtgata cctacatgat gggcaatcag caggggcagg gaacgggtat
     2041 cgataccggt atcaaacagt tcctgagtgg gctgggggcg ctgtcagcac agaccacgca
     2101 gctaccggca tttgatgccc tgcggcaggc gctgccgatg gtgcacagca ttatattgat
     2161 ggccctggtg atcagcattc cactggtgat gctgtttggt gcttacgacc cgaaggtggt
     2221 ggtcaccatt acgtttgcga tgtttgcgct gatctttgtt cccttctggt gggagttggg
     2281 cggctggctg gacgacaagc ttatccagtt gctgtacgcc agctacaagg ggtataccaa
     2341 cagcttcaac aactggattg ccagtaccgg ctcggatggc tggatcatga atctggtgct
     2401 gggggcgatg tatgtggtat tgccgatgtt ctggtttggg gcggtgagct ggagtggtgt
     2461 tcagatcgga ggagcattat cttcagcagt tgataatgcc agcaaaggtt cgcaacaggc
     2521 tggagctaaa ggacctggcg ttgcaggtga agtcgtcaaa acggtagcaa cgaaaggggc
     2581 atctaaaggg aaaggttgaa tcggtaccct cgagaagctt gcggccgcgg ggatccagac
     2641 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2701 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2761 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2821 gttttttcgg atcctctaga gtcgatctgc aggcatgcta gcttggcgta atcatggtca
     2881 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2941 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3001 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3061 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5041 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5101 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5161 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5221 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5281 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     5341 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5401 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     5461 tcccagtcac gacgttgtaa aacgacggcc agtgagttcg agcttgcatg cctgcaggtc
     5521 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5581 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5641 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5701 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5761 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5821 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5881 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5941 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     6001 cggtgggagg tctatataag cagagctcgt tt
//
