LOCUS       sequence-210847-        4856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_0557 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4856)
  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 4856)
  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_0557
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..172
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     164..181
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     165..181
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        208..511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     666..686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     712..736
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          853..949
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1010..1012,1013..1015,1016..1726)
                     /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(1055..1076)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1316..1335)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1663..1684
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2176..2310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2200..2219)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2254..2273
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2421..2437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2421..2437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2434..2456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2445..2461
                     /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(2469..2475,2476..2493,2494..2499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2514..2535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2652..2669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2823..3411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2903..2922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3582..4373,4374..4442))
                     /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     4205..4224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4443..4547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4615..4633
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4671..4693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4793..4812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
       61 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      121 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      181 tgagttcgag cttgcatgcc tgcaggtcgt tacataactt acggtaaatg gcccgcctgg
      241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      481 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      541 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      601 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      661 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      721 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      781 ccgggaccga tccagcctcc ggactctaga ggatccggta ctagaggaac tgaaaaacca
      841 gaaagttaac tggtaagttt agtctttttg tcttttattt caggtcccgg atccggtggt
      901 ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt tacttctagg cctgtacgga
      961 agtgttactt ctgctctaaa agctgcggaa ttgtacccgc gggcccacca tggtgagcaa
     1021 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1081 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1141 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1201 cttgacctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1261 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1321 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1381 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1441 caactacaac agccacaagg tctatatcac cgccgacaag cagaagaacg gcatcaaggt
     1501 gaacttcaag acccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1561 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1621 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1681 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagcgaa ttggtggagg
     1741 gggcggttcc ggtggcggag gttcccgaat tcgtcgacgg agatctgata tgaacaggaa
     1801 agaatatgct atgaaccctt atatttatct tggtggcgca atacttgcag aggtcattgg
     1861 tacaacctta atgaagtttt cagaggggtt tacacggtta tggccctctg ttggcacaat
     1921 tatttgctat tgtgcatcat tctggttatt agctcagacg ctggcttata tacctacagg
     1981 gattgcttat gctatctggt caggagtcgg tatcgttctg attagtttgc tttcctgggg
     2041 attttttggt caacgactgg acctgccagc tatcataggc atgatgttga tttgtgccgg
     2101 tgtgttagtc attaacttgt tgtcacgaag tgcaccgcat tgaatcggta ccctcgagaa
     2161 gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2221 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2281 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2341 caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat
     2401 gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2461 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2521 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2581 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2881 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2941 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3001 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3061 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3121 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3181 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3241 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3301 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3361 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3421 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3481 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4441 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4501 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4561 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4621 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4801 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgt
//
