LOCUS       sequence-210534-        4868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_004 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4868)
  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 4868)
  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_004
FEATURES             Location/Qualifiers
     source          1..4868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          131..227
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(288..290,291..293,294..1004)
                     /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(333..354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(594..613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     941..962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1466..1600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1490..1509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1544..1563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1711..1727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1711..1727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1724..1746)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1735..1751
                     /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(1759..1765,1766..1783,1784..1789))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1942..1959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2113..2701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2193..2212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2872..3663,3664..3732))
                     /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     3495..3514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3733..3837)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3905..3923
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3961..3983)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4083..4102
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4296..4318
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4310..4327
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4311..4327
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4354..4657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4658..4861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4812..4832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4858..4868,1..14)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
       61 gggaccgatc cagcctccgg actctagagg atccggtact agaggaactg aaaaaccaga
      121 aagttaactg gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg
      181 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag
      241 tgttacttct gctctaaaag ctgcggaatt gtacccgcgg gcccaccatg gtgagcaagg
      301 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      361 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      421 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      481 tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      541 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      601 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      661 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      721 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
      781 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      841 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      901 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      961 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcgaatt ggtggagggg
     1021 gcggttccgg tggcggaggt tcccgaattc gtcgacggag atctgatatg acgtatggag
     1081 aagcgtatct ggaagggtgg aaaaacatct ttaattatga aggtgtttcc aatcgcttcg
     1141 agttctggtc gtttatgatt ggtagtggcg ttatctgtct gcttccgctt ttatgttggt
     1201 ggctagcggt aacaattaac aatgactatg gtgttttcat tttttttgcg ctgcccgtgt
     1261 catttatttt aacgttgatt tttgccgtac cggcgatagc gttagccgtc agaaggatgc
     1321 acgatatcgg atattcaggg tggtgggtta ccattgccgt gttgatccct gttacgggcg
     1381 taattctgct tatactgtgc tgcctgccgt caaagtcaca agaccaggca tgaatcggta
     1441 ccctcgagaa gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg
     1501 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1561 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1621 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat
     1681 ctgcaggcat gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1741 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1801 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1861 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2521 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3841 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3901 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     3961 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4021 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4081 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4141 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     4201 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     4261 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     4321 ggccagtgag ttcgagcttg catgcctgca ggtcgttaca taacttacgg taaatggccc
     4381 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     4441 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     4501 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     4561 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     4621 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     4681 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     4741 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     4801 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     4861 tcgtttag
//
