LOCUS       sequence-210522-        4679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_779 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4679)
  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 4679)
  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_779
FEATURES             Location/Qualifiers
     source          1..4679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          134..230
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(291..293,294..296,297..1007)
                     /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(336..357)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(597..616)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     944..965
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1280..1414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1304..1323)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1358..1377
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1525..1541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1525..1541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1538..1560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1549..1565
                     /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(1573..1579,1580..1597,1598..1603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1618..1639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1756..1773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1927..2515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2007..2026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2686..3477,3478..3546))
                     /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     3309..3328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3547..3651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3719..3737
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3775..3797)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3897..3916
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4110..4132
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4124..4141
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4125..4141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4168..4471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4472..4675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4626..4646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4672..4679,1..17)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
       61 accgggaccg atccagcctc cggactctag aggatccggt actagaggaa ctgaaaaacc
      121 agaaagttaa ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg gatccggtgg
      181 tggtgcaaat caaagaactg ctcctcagtg gatgttgcct ttacttctag gcctgtacgg
      241 aagtgttact tctgctctaa aagctgcgga attgtacccg cgggcccacc atggtgagca
      301 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      361 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      421 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      481 ccttgaccta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      541 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      601 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      661 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      721 acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
      781 tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      841 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
      901 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      961 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagcga attggtggag
     1021 ggggcggttc cggtggcgga ggttcccgaa ttcgtcgacg gagatctgat atgttaaaac
     1081 atcgttatgc aaatacggaa gtgaaagtta ctcacagcac attgaataaa cggtatgatg
     1141 aagaaattgc aaacaacaca acaaggagcc acgcatggaa atgctcgaag agcaccgctg
     1201 ttttgaaggc tggcagcaac gctggcgaca cgactccagt accttgaatc ggtaccctcg
     1261 agaagcttgc ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     1321 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1381 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1441 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     1501 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1681 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2821 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3661 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3721 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3781 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3841 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3901 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     3961 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4021 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4081 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4141 tgagttcgag cttgcatgcc tgcaggtcgt tacataactt acggtaaatg gcccgcctgg
     4201 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4261 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4321 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4381 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4441 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4501 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4561 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4621 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgtt
//
