LOCUS       sequence-456140-        6236 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of human EHMT1 with deletion of N-terminal 300 aa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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_Ank+SET
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        306..609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        610..813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     764..784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      852..861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(858..860,861..863,864..1574)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(858..860,861..863,864..1574)
                     /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(903..924)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1164..1183)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1511..1532
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3269..3390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3306..3325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3360..3379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3397..3852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3534..3555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3746..3765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3879..3983
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3985..4342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3985..4176
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4008..4028)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4193..4328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4377..5171
                     /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             4377..5171
                     /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(4431..4450)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5041..5060
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5359..5378)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5403..5450
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5779..6236,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
      241 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      301 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      361 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      421 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      481 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      541 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      601 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      661 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      721 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      781 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      841 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      901 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      961 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1021 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1081 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1141 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1201 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1261 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1321 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1381 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1441 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1501 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1561 acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgaattcg ttccacccaa
     1621 agcagctgta cttctccgcc aggcaagggg agcttcagaa ggtgctcctc atgctggtgg
     1681 acggaattga ccccaacttc aaaatggagc accagaataa gcgctctcca ctgcacgccg
     1741 cggcagaggc tggacacgtg gacatctgcc acatgctggt tcaggcgggc gctaatattg
     1801 acacctgctc agaagaccag aggaccccgt tgatggaagc agccgaaaac aaccatctgg
     1861 aagcagtgaa gtacctcatc aaggctgggg ccctggtgga tcccaaggac gcagagggct
     1921 ctacgtgttt gcacctggct gccaagaaag gccactacga agtggtccag tacctgcttt
     1981 caaatggaca gatggacgtc aactgtcagg atgacggagg ctggacaccc atgatctggg
     2041 ccacagagta caagcacgtg gacctcgtga agctgctgct gtccaagggc tctgacatca
     2101 acatccgaga caacgaggag aacatttgcc tgcactgggc ggcgttctcc ggctgcgtgg
     2161 acatagccga gatcctgctg gctgccaagt gcgacctcca cgccgtgaac atccacggag
     2221 actcgccact gcacattgcc gcccgggaga accgctacga ctgtgtcgtc ctctttcttt
     2281 ctcgggattc agatgtcacc ttaaagaaca aggaaggaga gacgcccctg cagtgtgcga
     2341 gcctcaactc tcaggtgtgg agcgctctgc agatgagcaa ggctctgcag gactcggccc
     2401 ccgacaggcc cagccccgtg gagaggatag tgagcaggga catcgctcga ggctacgagc
     2461 gcatccccat cccctgtgtc aacgccgtgg acagcgagcc atgccccagc aactacaagt
     2521 acgtctctca gaactgcgtg acgtccccca tgaacatcga cagaaatatc actcatctgc
     2581 agtactgcgt gtgcatcgac gactgctcct ccagcaactg catgtgcggc cagctcagca
     2641 tgcgctgctg gtacgacaag gatggccggc tcctgccaga gttcaacatg gcggagcctc
     2701 ccttgatctt cgaatgcaac cacgcgtgct cctgctggag gaactgccga aatcgcgtcg
     2761 tacagaatgg tctcagggca aggctgcagc tctaccggac gcgggacatg ggctggggcg
     2821 tgcggtccct gcaggacatc ccaccaggca cctttgtctg cgagacgggg aggtttactg
     2881 catcgacgcg cggttctacg ggaacgtcag ccggttcatc aaccaccact gcgagcccaa
     2941 cctggtgccc gtgcgcgtgt tcatggccca ccaggacctg cggttccccc ggatcgcctt
     3001 cttcagcacc cgcctgatcg aggccggcga gcagctcggg tttgactatg gagagcgctt
     3061 ctgggacatc aaaggcaagc tcttcagctg ccgctgcggc tcccccaagt gccggcacgt
     3121 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     3181 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3241 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3361 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     3421 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3481 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3541 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3601 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3661 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3721 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3781 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3841 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3901 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3961 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4021 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4081 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4141 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4201 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4261 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4321 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4381 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4441 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4501 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4561 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4621 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4681 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4741 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4801 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4861 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4921 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4981 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5041 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5101 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5161 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5221 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5281 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5341 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5401 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5461 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5521 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5581 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5881 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcg
//
