LOCUS       sequence-210465-        5372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_701 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5372)
  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 5372)
  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_701
FEATURES             Location/Qualifiers
     source          1..5372
                     /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"
     CDS             1543..1554
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1970..2104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1994..2013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2048..2067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2215..2231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2215..2231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2228..2250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2239..2255
                     /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(2263..2269,2270..2287,2288..2293))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2308..2329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2446..2463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2617..3205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2697..2716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3376..4167,4168..4236))
                     /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     3999..4018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4237..4341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4409..4427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4465..4487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4587..4606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4800..4822
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4814..4831
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4815..4831
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4858..5161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5162..5365
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5316..5336
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5362..5372,1..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 gatgtattca
     1081 aaatttccgt tgatacacag caggttcaca agatcactca ggagaacttt gatctcgcaa
     1141 ctaaaaagca ggcaccctgg tatcagccaa tcaatggtca ccggggatgg tttgcggttt
     1201 gtccggcttg tgacaacccg atccagatta ttggaatgct tgaacgtgat gcgccgtatg
     1261 ggaagcactt cttcccgact gccggtgcag tggctgtacc gttgaaaggg cgtgttgata
     1321 aggaggaata tgagtggtgc ccatatgcca gcaagaataa gagtctaacg aaagatgctc
     1381 ggcgagcaga gggtagtgaa cttgcgctat tgataaaaaa aaccttaatt gagcaatttg
     1441 accgggttat ttatttgtta gaaaagggca tcgggatgag aatttcgggg gccttggcga
     1501 aacagatgct ggaagattac cggctggctg aaggatggcg ttatagaggg gcgacaagac
     1561 agaatatccc ttgggtgttc gcttatatga tgcaggctaa gtgggtgaaa gggcgcatat
     1621 tctacgacaa aaatattctg gaagaacttt gtaccaaacg aaccgattta atttttaacg
     1681 acaaagggaa acttactact aaagatgaaa aaaaattttg cgacttaagt tttagtttca
     1741 tcaaacaccg tcgccatgta gagcagcata ctttgtctga aagtatcgag ttcaatatcg
     1801 caaactcaaa acaggaaacg gtatacaaaa agactattgc tttcgaatat ggcgatttcc
     1861 tgaggctgat aaacagtaaa aatgaggctt acagaaaaat ggaccttgtt gatcttgcca
     1921 gaagcgtctt ggcctgaatc ggtaccctcg agaagcttgc ggccgcgggg atccagacat
     1981 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2041 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2101 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2161 tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata
     2221 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2281 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2341 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4381 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4561 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4621 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     4681 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     4741 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     4801 ccagtcacga cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc tgcaggtcgt
     4861 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4921 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4981 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5041 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5101 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     5161 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     5221 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     5281 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     5341 gtgggaggtc tatataagca gagctcgttt ag
//
