LOCUS       sequence-456138-        5586 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of human EHMT1 N-terminal 300 aa only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  AUTHORS   Biligiri KK, Sharma NR, Mohanty A, Sarkar DP, Vemula PK, Rampalli S.
  TITLE     A cytoplasmic form of EHMT1N methylates viral proteins to enable 
            inclusion body maturation and efficient viral replication
  JOURNAL   PLOS Biology 22(11): e3002871.
REFERENCE   2  (bases 1 to 5586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLOS 
            Biology 22(11): e3002871."
COMMENT     Sequence Label: EGFP_EHMT1_N300
FEATURES             Location/Qualifiers
     source          1..5586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    353..400
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     425..444
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(632..1426)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(632..1426)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(743..762)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1353..1372
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1461..1818)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1475..1610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1627..1818
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1775..1795
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1951..2406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2038..2057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2248..2269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2413..2534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2424..2443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2478..2497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3579..4289,4290..4292,4293..4295))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3579..4289,4290..4292,4293..4295))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3621..3642)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3970..3989
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4229..4250
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4292..4301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4340..4543)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4369..4389)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4544..4847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(5022..5586,1..24))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5102..5121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
       61 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      121 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      181 cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc
      241 gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt
      301 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
      361 gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
      421 ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc
      481 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
      541 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
      601 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
      661 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
      721 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
      781 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
      841 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
      901 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
      961 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     1021 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     1081 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     1141 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     1201 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     1261 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     1321 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     1381 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     1441 atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     1501 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     1561 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     1621 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     1681 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     1741 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
     1801 tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     1981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2161 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2281 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2341 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     2401 caatttacgc gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2461 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2521 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2581 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     2641 agttatctag atccggtgga tcccgggccc gcggtaccgt cgacaggaac caggctatag
     2701 gttcccattc ttcgtttttt cttccgaggt actgcagctg ctgtctgtga ttttgtggtg
     2761 gccatgtagc actggttctg aggtagcgac tgatgaaggg atgggaaggg gggtaaaagc
     2821 tgctgtcgtc caaagtcaga aatgtttttg ttgacctcct cttttggctc cttatgatct
     2881 ctagcttctc gaacttctct gggatcttta ctggctgcat gcaggcccac gacggacttc
     2941 ggcatggtct tgcgtgccct gtggaccttg acgtcggcgc caggggccgg gagcttcctg
     3001 tcctctgtgt cagcactccc ctccccaagg gtggctggtg gggcggctgg cgtctgggga
     3061 aaagcgcttg gagtcctgcc tttgccagcc cctccaggaa gggtttttgc agcatggcca
     3121 ggcagcgaag aggccagagt gctggtagtc ctcaagggct gtgcctgtag ggccggctta
     3181 tttaagatgt atccgttgct gccgatgaca gaagtctgca caaagtcgtc ggcagtgacg
     3241 tggttttgct tcgccgcttc tgagtctctt tctgaaaccc cattttccgc tatccgagtt
     3301 agtgtgttgg tgccatcctg ggggttgacc cttgcgctgt cctgagtgtg ctttgcagca
     3361 tttgcatgac tgctggcatc gctgttttca caagacccat aggtctcacc gtccgcagtc
     3421 atgtgggcct ctcctgcctg tttctctgct gagccttcgt cggcagccat aggtgtctct
     3481 tctcccagca gctcggtttt cacacagcaa tcctgctgag gctcccccct cgccggaact
     3541 gcctcggcat cggcggcggc catggatctg agtccggact tgtacagctc gtccatgccg
     3601 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     3661 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     3721 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     3781 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     3841 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     3901 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     3961 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     4021 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     4081 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     4141 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     4201 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     4261 accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcgga
     4321 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     4381 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     4441 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     4501 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     4561 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     4621 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     4681 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     4741 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     4801 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     4861 atgggctatg aactaatgac cccgtaattg attactatta ataactaatg catggcggta
     4921 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5581 cagatt
//
