LOCUS       sequence-210218-        5099 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_671 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5099)
  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 5099)
  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_671
FEATURES             Location/Qualifiers
     source          1..5099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..116
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     108..125
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     109..125
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        152..455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        456..659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     610..630
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     656..680
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          797..893
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(954..956,957..959,960..1670)
                     /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(999..1020)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1260..1279)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1607..1628
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2363..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2608..2624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2608..2624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2621..2643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2632..2648
                     /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(2656..2662,2663..2680,2681..2686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2701..2722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2839..2856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3769..4560,4561..4629))
                     /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     4392..4411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4630..4734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4802..4820
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4858..4880)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4980..4999
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
       61 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
      121 ccagtgagtt cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc
      181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      661 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
      721 gacaccggga ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa
      781 accagaaagt taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg
      841 tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta
      901 cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccgcgggccc accatggtga
      961 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1021 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1081 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1141 ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1201 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1261 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1321 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1381 agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca
     1441 aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1501 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1561 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1621 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag cgaattggtg
     1681 gagggggcgg ttccggtggc ggaggttccc gaattcgtcg acggagatct gatatgaacc
     1741 ttcatatccc aatggctgtc tgcctgacgg ccagcatcct ggccggttgt acccaaaagc
     1801 tccagcaacc tgatacgcaa cctcatccat cggcgctaaa tggcaacgtc cagccagccc
     1861 gactggatgt ctatgccgcc tctcctgaag ttgttcgcta tgaccgctat ctgctggttg
     1921 atacttcccc gagcgccgca cagcgtgctc cgttggaaca gaccattgat atccgtatcc
     1981 ccgcgtcatt atcgccgacc gtggcggatg ccatgcgcta tgccctgcgc cagtccgggt
     2041 actcgttgtg cagtacgaca ccggcgaacc gggtgctgta caaccagacg ttgcctgccg
     2101 tgcattatca actggggcca atgcggctga gtcaggcatt acaaattctg gccggatcgg
     2161 cctggcagat cgaggttgat tctgtacaac ggatcgtctg ccactcattg cgtgatggct
     2221 atcgtttgcc gccggaagcg gtgaaaacgt cggtcacttc ggccactgtg ccgctacccg
     2281 ctcaacccgt caatatcacc gttccggctc cgcagaggat gctgaaatga atcggtaccc
     2341 tcgagaagct tgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca
     2401 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgatctg
     2581 caggcatgct agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4741 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4801 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4861 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4921 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4981 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5041 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggg
//
