LOCUS       sequence-209620-        5774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_130 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5774)
  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 5774)
  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_130
FEATURES             Location/Qualifiers
     source          1..5774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        44..347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        348..551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     502..522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     548..572
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          689..785
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(846..848,849..851,852..1562)
                     /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(891..912)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1152..1171)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1499..1520
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2930..3064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2954..2973)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3008..3027
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3175..3191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3175..3191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3188..3210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3199..3215
                     /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(3223..3229,3230..3247,3248..3253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3268..3289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3406..3423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3577..4165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3657..3676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4336..5127,5128..5196))
                     /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     4959..4978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5197..5301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5369..5387
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5425..5447)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5547..5566
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5760..5774,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5774,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca taacttacgg
       61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      541 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      601 acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat ccggtactag
      661 aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg
      721 tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact
      781 tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt acccgcgggc
      841 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      901 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      961 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1021 ccaccctcgt gaccaccttg acctacggcg tgcagtgctt cgcccgctac cccgaccaca
     1081 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1141 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1201 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1261 ggcacaagct ggagtacaac tacaacagcc acaaggtcta tatcaccgcc gacaagcaga
     1321 agaacggcat caaggtgaac ttcaagaccc gccacaacat cgaggacggc agcgtgcagc
     1381 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1441 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1501 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1561 agcgaattgg tggagggggc ggttccggtg gcggaggttc ccgaattcgt cgacggagat
     1621 ctgatatgcc taacaatctc gacagtaacg tcagtcaaat cgttctgaaa aaattccttc
     1681 ctggttttat gtcagattta gttctggcga aaaccgtaga ccgtcagttg ctggcaggtg
     1741 aaatcaactc cagcactggc gatagcgtta gctttaaacg tccgcatcaa ttctcatccc
     1801 tccgtactcc cactggtgat atttcaggga aaaataaaaa caacctgatc tcaggtaaag
     1861 ctacggggcg tgtaggtaat tacatcactg ttgctgttga atatcagcaa ctggaggaag
     1921 cgatcaagct taaccaactg gaagaaattc tcgcgccggt tcgccagcga atcgttaccg
     1981 accttgaaac agagcttgct cacttcatga tgaataacgg tgcgttgtca cttggtagcc
     2041 ccaatactcc aatcaccaaa tggtctgatg ttgcgcagac ggcatctttc ctgaaagacc
     2101 tcggcgttaa tgaaggtgaa aactatgctg taatggatcc atggtctgca cagcgacttg
     2161 ctgatgcgca gactggtttg cacgcttcag atcaattggt tcgtactgca tgggagaatg
     2221 cgcagattcc aaccaatttt ggcggcattc gcgcactgat gtctaatggg cttacctctc
     2281 gtacgcaggg ggcatttggc ggaacactga cagtcaaaac acagccaact gttacctata
     2341 acgcagttaa agactcatac cagttcactg taacattgac cggagcgaca gccagcgtta
     2401 caggttttct gaaagctggt gatcaggtta aattcaccaa tacctactgg ctgcaacagc
     2461 agaccaaaca ggcgttgtat aacggagcca caccaattag cttcactgca acggttactg
     2521 ctgatgctaa ttcagacagc agtggcgatg tgacggttac gctttctggt gttccgattt
     2581 atgacactac aaacccgcag tacaactctg taagtcgtca ggtagcggca gacgatgccg
     2641 tatctgtagt aggcactgct agccagacaa tgaagccaaa cctgttctat aacaagttct
     2701 tctgtggact tggctctatc ccactgccga aactgcacag tattgattct gctgttgcaa
     2761 catatgaagg tttctccatc cgcgtacata aatacgcaga tggcgatgcc aacgtgcaaa
     2821 aaatgcgctt tgacttactg cctgcatatg tgtgctttaa ccctcacatg ggcggtcagt
     2881 tcttcggtaa tccgtgaatc ggtaccctcg agaagcttgc ggccgcgggg atccagacat
     2941 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3001 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3061 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3121 tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata
     3181 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3241 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3301 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3361 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5341 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5401 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5461 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5521 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5581 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5641 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5701 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5761 ccagtcacga cgtt
//
