LOCUS       sequence-212746-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_806 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  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 4784)
  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_806
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     96..113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     97..113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        140..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     644..668
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          785..881
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(942..944,945..947,948..1658)
                     /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(987..1008)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1248..1267)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1595..1616
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2036..2170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2114..2133
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2281..2297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2281..2297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2294..2316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2305..2321
                     /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(2329..2335,2336..2353,2354..2359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2374..2395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2512..2529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2683..3271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2763..2782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3442..4233,4234..4302))
                     /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     4065..4084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4303..4407)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4475..4493
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4531..4553)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4653..4672
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
       61 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagttcg
      121 agcttgcatg cctgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      661 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      721 gatccagcct ccggactcta gaggatccgg tactagagga actgaaaaac cagaaagtta
      781 actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa
      841 tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg gaagtgttac
      901 ttctgctcta aaagctgcgg aattgtaccc gcgggcccac catggtgagc aagggcgagg
      961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1081 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttgacct
     1141 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1381 acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca
     1441 agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1561 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1621 ccgccgggat cactctcggc atggacgagc tgtacaagcg aattggtgga gggggcggtt
     1681 ccggtggcgg aggttcccga attcgtcgac ggagatctga tatgactaat cgcttatctc
     1741 tggcatttat gcctgtttct atcacgcttc cggcttggga acatgccgtt gaagtatttg
     1801 atttctccca gtgggaacgt cggcaatttg cgctgatcaa agccgcgcaa gatgcctgga
     1861 accatcgttc agatcctgat actcagcagg tgacattttc gctgacgttg tttgttcgtc
     1921 tgggaggtga aactaccgaa cggacgcaaa actttgtcgc cagatatgtt gacgatgcac
     1981 tggttgtaac cctgggagaa tgaatcggta ccctcgagaa gcttgcggcc gcggggatcc
     2041 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2101 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2161 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2221 ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg cgtaatcatg
     2281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4441 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4501 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4561 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4621 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4681 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     4741 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcgg
//
