LOCUS       sequence-210852-        5753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_604 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5753)
  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 5753)
  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_604
FEATURES             Location/Qualifiers
     source          1..5753
                     /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    3073..3207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3097..3116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3151..3170
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3318..3334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3331..3353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3342..3358
                     /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(3366..3372,3373..3390,3391..3396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3411..3432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3549..3566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3720..4308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3800..3819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4479..5270,5271..5339))
                     /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     5102..5121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5340..5444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5512..5530
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5568..5590)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5690..5709
                     /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 tggcaaagcc
     1801 gatgaataat gaatcgattt atcagcaata tatgtacagt tacccgcata agaccgctta
     1861 tcgggaatta aataatattg cttttgcgga tgtgaagtcg cgtatttacg agcacaaaac
     1921 acatctctat attcatatgc ctttttgcca gagccgctgc gggttttgta atctctttac
     1981 ctgcacgggg gcagacaatg cctttatcga caactacata gaggcaatta tcgttcaggc
     2041 ccagcaaatg caacttgaag ccgtagactg ggcaagtttt accattggcg gcggtacgcc
     2101 attgttgctt aatgtcgggc agctgtcacg gttgtttcgc ggggtatttg acgaattaca
     2161 ggtcgatgcg catattttta aggcaattga aacatcgcct tccgatacca ccgccgaaaa
     2221 agtcgcgtta ttgcgccagc ttgcggttaa ccgtgtcagt attggtgtgc agagttttaa
     2281 cgatctggaa cttgccacat tacaccgccg ccatagtgcg gaaaatgccc atcgagcgct
     2341 ggagctactg cacaggggta atttcccgat aatcaatatt gatattattt acggtattcc
     2401 cggtcaaacg gcagcaagtt taatggcgtc gcttaatcag gcattagatt atcaacctga
     2461 cgaactgttt ctgtacccgc tgtataatat gccgcgtcag ccggatatgt ataacctcta
     2521 tattgaagcc cgcgatcgct tattacatga atgttatacg caaacgtcaa tgcgccgttt
     2581 tgtgcgggtg aataatgatt cagcggctga aagttgtggc tttgaaaatt cgctggcgct
     2641 tggcgcgggc ggacgcagtt atcttggcaa tctacattac tgcacgccgt ggtcgcaaaa
     2701 ccagcctgca tcgcggcaaa ttgtcgcgga ttttatcgac cgaccagata agacccttat
     2761 tagccacggc tatgtgcttt cacaagcaga gatgaagcgg cgttttatta tcaagaatct
     2821 ctttttctat aaagggcttt cgttaagtga ttatcagcac cagttctcct cacatgcatt
     2881 aagcgatttt cctctgctgg gagatttcat taaacaagat tattgccacc agcaggataa
     2941 tcgcattctg ctcaccgcgc agggtttagc gctttcggac cagcttggcc cgctgtttat
     3001 ttccccggaa gttcgcaggc tggagacgca taaacgatga atcggtaccc tcgagaagct
     3061 tgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     3121 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3181 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3241 gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgatctg caggcatgct
     3301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5461 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5521 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5581 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5641 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     5701 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgt
//
