LOCUS       sequence-210249-        5375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_605 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5375)
  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 5375)
  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_605
FEATURES             Location/Qualifiers
     source          1..5375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(61..63,64..66,67..777)
                     /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(106..127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(367..386)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     714..735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1746..1880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1770..1789)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1824..1843
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1991..2007)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1991..2007)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2004..2026)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2015..2031
                     /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(2039..2045,2046..2063,2064..2069))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2084..2105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2222..2239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2393..2981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2473..2492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3152..3943,3944..4012))
                     /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     3775..3794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4013..4117)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4185..4203
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4241..4263)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4363..4382
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4576..4598
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4590..4607
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4591..4607
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4634..4937
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4938..5141
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5092..5112
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5138..5162
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5279..5375
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 gcctgtacgg aagtgttact tctgctctaa aagctgcgga attgtacccg cgggcccacc
       61 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      121 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      181 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      241 ctcgtgacca ccttgaccta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      301 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      361 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      421 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      481 aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac
      541 ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc
      601 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      661 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      721 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagcga
      781 attggtggag ggggcggttc cggtggcgga ggttcccgaa ttcgtcgacg gagatctgat
      841 atgtataacg taacagacga agtaccggaa aatacccgca cggcaatcca ttcctgcgat
      901 ggcgaaatca agatgaatga tttgatagct agggaggcga atgttctgtt tgtagtgctt
      961 gattccctgc gttttgatat tgcggttcag gaacaagagg cgggcaatac gccgaacatt
     1021 aaccgctatg gtcgctggat caagtgcgaa gccgctggca attttacctg gccttcgcat
     1081 cattcgatgt tctctggttt tatgcctaag ccgctggaag cggcaccggg tcattcaatg
     1141 ctctttttcc cgaaagatat tggtttaggg cgtttagggc caaaaaatgc gttcgcattc
     1201 agcgaagaga cgtggattaa agggctggaa aacaaaggtt accgcactat ctgcgtgggt
     1261 ggcgtctcct tttttaataa ccgttcaggg atggggaaag tgttcccggc gatgtttaaa
     1321 gagagttact ggtatcctaa tttctcctgc acggtaaaag agagcatgga taatcaactc
     1381 accttaatta ataaaaaggt aaatgctgaa catccgctga tgatgtatat caatatcgac
     1441 accattcatt atcccaatca tttttatatt gaaaatgcca agcctggcga tacggtagaa
     1501 actcacgccg ctgccctgca ttatattgat gcgagaatcg aaaaactttt cgatatcttt
     1561 cggcaaaccg ggcgggagac gttagttatt ctctgctccg atcatggtac ctgctacggg
     1621 gaagacggta aatatttcca cagttttaac catccgattg tcaataccgt accctatttt
     1681 cactttgtgt tagatggcaa agccgatgaa tgaatcggta ccctcgagaa gcttgcggcc
     1741 gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1801 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1861 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1921 gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg
     1981 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2041 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2101 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2161 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2221 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2281 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2341 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2401 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2461 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2521 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2581 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2641 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2701 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2761 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2821 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2881 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3061 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3301 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3361 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3421 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3481 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3541 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3601 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3661 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3721 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3781 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3841 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3901 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3961 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4141 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4201 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4261 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4321 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4381 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4441 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4501 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4561 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
     4621 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4681 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4741 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4801 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4861 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4921 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4981 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5041 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5101 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     5161 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
     5221 gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa gttaactggt
     5281 aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag
     5341 aactgctcct cagtggatgt tgcctttact tctag
//
