LOCUS       sequence-210660-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_312 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  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 4835)
  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_312
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     240..262
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     254..271
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     255..271
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        298..601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        602..805
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     756..776
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     802..826
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          943..1039
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1100..1102,1103..1105,1106..1816)
                     /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(1145..1166)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1406..1425)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1753..1774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2245..2379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2269..2288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2323..2342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2490..2506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2490..2506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2503..2525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2514..2530
                     /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(2538..2544,2545..2562,2563..2568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2583..2604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2721..2738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2892..3480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2972..2991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3651..4442,4443..4511))
                     /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     4274..4293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4512..4616)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4684..4702
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4740..4762)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
       61 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      121 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      181 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      241 ccagtcacga cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc tgcaggtcgt
      301 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      361 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      421 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      481 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      541 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      601 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      661 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      721 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      781 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      841 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctaga
      901 ggatccggta ctagaggaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg
      961 tcttttattt caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg
     1021 atgttgcctt tacttctagg cctgtacgga agtgttactt ctgctctaaa agctgcggaa
     1081 ttgtacccgc gggcccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1141 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1201 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1261 gcccgtgccc tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt gcttcgcccg
     1321 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1381 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1441 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1501 cggcaacatc ctggggcaca agctggagta caactacaac agccacaagg tctatatcac
     1561 cgccgacaag cagaagaacg gcatcaaggt gaacttcaag acccgccaca acatcgagga
     1621 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1681 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     1741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1801 ggacgagctg tacaagcgaa ttggtggagg gggcggttcc ggtggcggag gttcccgaat
     1861 tcgtcgacgg agatctgata tggacagagg ggaaatctgg ctggtttcac tggatccgat
     1921 tgcaggtcac gagcaaagcg gaaaacgtcc ggtgcttatc gtatcgaagg catcgtttaa
     1981 caagctgacc cggctacccg ttgttgttcc cgttaccagc ggcggaaact tcgcccgtgc
     2041 agccggtttt accgtctcac tggacggtgc gggaacaaaa acaacgggcg ttattcgctg
     2101 cgaccagccc agaaccattg atatgggagc ccggagcggt aagcgactgg aacgtatacc
     2161 cgatgctgtc gtgaatgaag tgctggcccg actggaggcc attctgagct gaatcggtac
     2221 cctcgagaag cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga
     2281 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2341 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2401 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc
     2461 tgcaggcatg ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2521 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2581 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2641 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3601 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3661 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3721 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3781 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3841 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3901 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3961 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4021 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4081 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4141 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4201 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4261 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4321 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4381 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4441 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4501 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4561 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4621 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4681 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4741 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4801 agacaagccc gtcagggcgc gtcagcgggt gttgg
//
