LOCUS       sequence-210451-        4904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_687 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4904)
  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 4904)
  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_687
FEATURES             Location/Qualifiers
     source          1..4904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          137..233
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(294..296,297..299,300..1010)
                     /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(339..360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(600..619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     947..968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1508..1642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1532..1551)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1586..1605
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1753..1769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1753..1769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1766..1788)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1777..1793
                     /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(1801..1807,1808..1825,1826..1831))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1846..1867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1984..2001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2155..2743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2235..2254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2914..3705,3706..3774))
                     /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     3537..3556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3775..3879)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3947..3965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4003..4025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4125..4144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4338..4360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4352..4369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4353..4369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4396..4699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4700..4903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4854..4874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4900..4904,1..20)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
       61 gacaccggga ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa
      121 accagaaagt taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg
      181 tggtggtgca aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta
      241 cggaagtgtt acttctgctc taaaagctgc ggaattgtac ccgcgggccc accatggtga
      301 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      361 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      421 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      481 ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      541 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      601 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      661 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      721 agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca
      781 aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
      841 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      901 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      961 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag cgaattggtg
     1021 gagggggcgg ttccggtggc ggaggttccc gaattcgtcg acggagatct gatatgaaag
     1081 gatacggttt attcatgctg gtcgcgttgt tgctgagcgg ttgcagtacc tccaaagatg
     1141 aaatgctgcc acccggtgac agtaccatgc tcgatatgtg gcagaacaaa gccagcagcg
     1201 cccggaccac gacagaagcg cgtagcgtgt tacgccgggg gctgacggat tcggatctgg
     1261 caacccgtca gtcactggaa gagagctata cccgcgatgc tcagaatgag atccagcagc
     1321 ttttcccgcg cctgcctaac ccggatatgg tgatttatat ctatccgcat atggcaggaa
     1381 atgagcctgc gccggtgccg ggatacagct cggtcttctc tttctatagc cgggtacagt
     1441 atgcattacc gggcgaacgg atggaggcac tctgaatcgg taccctcgag aagcttgcgg
     1501 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1561 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1621 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1681 gggggaggtg tgggaggttt tttcggatcc tctagagtcg atctgcaggc atgctagctt
     1741 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1801 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1861 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1921 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1981 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2041 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2881 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2941 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3001 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3061 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3121 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3181 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3241 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3301 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3361 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3421 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3481 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3541 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3601 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3661 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3721 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3781 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3841 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3901 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3961 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4021 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4081 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4141 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4201 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4261 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4321 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agttcgagct
     4381 tgcatgcctg caggtcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4441 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4501 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4561 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4621 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     4681 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     4741 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     4801 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     4861 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctc
//
