LOCUS       sequence-210161-        4880 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_659 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  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 4880)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_659
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     585..609
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          726..822
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(883..885,886..888,889..1599)
                     /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(928..949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1189..1208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1536..1557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2073..2207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2097..2116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2151..2170
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2318..2334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2331..2353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2342..2358
                     /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(2366..2372,2373..2390,2391..2396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2411..2432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2549..2566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2720..3308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2800..2819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3479..4270,4271..4339))
                     /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     4102..4121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4340..4444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4512..4530
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4568..4590)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4690..4709
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagttc
       61 gagcttgcat gcctgcaggt cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
      601 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
      661 cgatccagcc tccggactct agaggatccg gtactagagg aactgaaaaa ccagaaagtt
      721 aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa
      781 atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac ggaagtgtta
      841 cttctgctct aaaagctgcg gaattgtacc cgcgggccca ccatggtgag caagggcgag
      901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttgacc
     1081 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1321 aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc
     1381 aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1501 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1561 gccgccggga tcactctcgg catggacgag ctgtacaagc gaattggtgg agggggcggt
     1621 tccggtggcg gaggttcccg aattcgtcga cggagatctg atatgaaact ctatcgtgca
     1681 acgtacagtc tcttaacccg cgatggtcgg gtcacagttg atgagcagtg ggtctttttt
     1741 gaagccgaaa cctactctga cgcgctggag aaaatacccg ggctattgat gacgatgtgg
     1801 gattgcggtg agtcccaact ggaggtctgg aacctgatgc aggataccca gcttgttgat
     1861 ctcagcatgg atatggaggt tgattcccct cgggactggc gtttctttga ggttggttat
     1921 tccgctggtt ctgccgttta tcttgatgtc gaacgtgatg gctggccgtt gttccttgtc
     1981 tctccgtcaa atctggcccg tctgaataat gccttggtcg agtgccggag gccagcatga
     2041 atcggtaccc tcgagaagct tgcggccgcg gggatccaga catgataaga tacattgatg
     2101 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2161 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2221 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttcg gatcctctag
     2281 agtcgatctg caggcatgct agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3241 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4441 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4501 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4561 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4621 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4681 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     4741 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     4801 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     4861 gatgtgctgc aaggcgatta
//
