LOCUS       sequence-212760-        4841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_786 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4841)
  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 4841)
  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_786
FEATURES             Location/Qualifiers
     source          1..4841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     119..136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     120..136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        163..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..691
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          808..904
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(965..967,968..970,971..1681)
                     /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(1010..1031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1271..1290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1618..1639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2116..2250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2140..2159)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2194..2213
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2361..2377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2361..2377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2374..2396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2385..2401
                     /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(2409..2415,2416..2433,2434..2439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2454..2475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2592..2609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2763..3351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2843..2862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3522..4313,4314..4382))
                     /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     4145..4164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4383..4487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4555..4573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4611..4633)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4733..4752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
       61 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      121 taaaacgacg gccagtgagt tcgagcttgc atgcctgcag gtcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      721 cctccataga agacaccggg accgatccag cctccggact ctagaggatc cggtactaga
      781 ggaactgaaa aaccagaaag ttaactggta agtttagtct ttttgtcttt tatttcaggt
      841 cccggatccg gtggtggtgc aaatcaaaga actgctcctc agtggatgtt gcctttactt
      901 ctaggcctgt acggaagtgt tacttctgct ctaaaagctg cggaattgta cccgcgggcc
      961 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1021 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1141 caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1381 gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg acaagcagaa
     1441 gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca gcgtgcagct
     1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1561 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1681 gcgaattggt ggagggggcg gttccggtgg cggaggttcc cgaattcgtc gacggagatc
     1741 tgatatgcct gctgacattg ttgagataag tacagactta tacaatgaat taattgccgg
     1801 acagcaggaa ggggggaaat taatcacgtc agatgaaaat ggtttaccgg tactgaaatc
     1861 tccggcgatt gattatgtcg cacgtgctga aaatcagcga atgcagttac ttgctcatgc
     1921 cgataatgtc acagctgact ggcgggtgga attaatgctt ggtgatatca gcagtacaga
     1981 taaagaaaaa ctatctgcct ggatggacta caaaaaagaa gtaaaagccg tcgacacttc
     2041 gacggctcct gagattagct ggcctgagtt accggaggtg tgaatcggta ccctcgagaa
     2101 gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2161 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2221 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2281 caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat
     2341 gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2401 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2461 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4501 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4561 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4621 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4681 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4741 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4801 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc g
//
