LOCUS       sequence-210322-        5027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_663 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  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 5027)
  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_663
FEATURES             Location/Qualifiers
     source          1..5027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          124..220
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(281..283,284..286,287..997)
                     /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(326..347)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(587..606)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     934..955
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1618..1752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1642..1661)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1696..1715
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1863..1879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1876..1898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1887..1903
                     /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(1911..1917,1918..1935,1936..1941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1956..1977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2094..2111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3024..3815,3816..3884))
                     /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     3647..3666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3885..3989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4057..4075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4113..4135)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4235..4254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4448..4470
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4462..4479
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4463..4479
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4506..4809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4810..5013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4964..4984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5010..5027,1..7)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
       61 atccagcctc cggactctag aggatccggt actagaggaa ctgaaaaacc agaaagttaa
      121 ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat
      181 caaagaactg ctcctcagtg gatgttgcct ttacttctag gcctgtacgg aagtgttact
      241 tctgctctaa aagctgcgga attgtacccg cgggcccacc atggtgagca agggcgagga
      301 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      361 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      421 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttgaccta
      481 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      541 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      601 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      661 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      721 cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa
      781 gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
      841 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
      901 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
      961 cgccgggatc actctcggca tggacgagct gtacaagcga attggtggag ggggcggttc
     1021 cggtggcgga ggttcccgaa ttcgtcgacg gagatctgat atgaacggac tgttaacgca
     1081 gattgttatg gaactgaaat ccggcaatat ccgccgttgt gaggcgatgg ggttaaccct
     1141 ggaagagatt caggaactta acagcttaac cgttgaagat ctgcattatc tggtgaatac
     1201 gcaggtatcg cttctgacct tccgcatcaa ccataccaac ctgaacacca tgctggcgca
     1261 ggcccgtcgt gagcagaccc ggacacagcg tatcgacaga gcgttaacgc tgggtggctc
     1321 tatcgagctg atgcagcatt ttttcggcct gacggcgacg gaagtcagtt cccggcgacg
     1381 cctggtcggt gttgtgaccc ggcaggggcg cagccagatc cctacagaag agcaggaact
     1441 ggcggtgtgg cagcagtgga aggcatcgaa cgttgacaac atggaatcac tggatgcgct
     1501 ggaagccatg atgttaattg ctgagcagca ggacatttcg ttaaccgcag tctggacgtt
     1561 aaccaaagga tggagtgccg cctgaatcgg taccctcgag aagcttgcgg ccgcggggat
     1621 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1681 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1741 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1801 tgggaggttt tttcggatcc tctagagtcg atctgcaggc atgctagctt ggcgtaatca
     1861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4021 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4081 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4141 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4201 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4261 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4321 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4381 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4441 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agttcgagct tgcatgcctg
     4501 caggtcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4561 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4621 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4681 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4741 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4801 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4861 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4921 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4981 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccg
//
