LOCUS       Exported                7988 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of human DNMT3B1 with myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3/Myc-DNMT3B1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7988)
  AUTHORS   Chen ZX, Mann JR, Hsieh CL, Riggs AD, Chedin F
  TITLE     Physical and functional interactions between the human DNMT3L 
            protein and members of the de novo methyltransferase family.
  JOURNAL   J Cell Biochem. 2005 Aug 1;95(5):902-17.
  PUBMED    15861382
REFERENCE   2  (bases 1 to 7988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7988
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      897..906
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             906..935
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3543..3561)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3544..3561)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3581..3598)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3587..3811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3857..4285
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3944..3963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4154..4175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4294..4314)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4299..4628
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4479..4614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4541..4560
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4695..5489
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4749..4768)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5359..5378
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5663..5784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5700..5719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5754..5773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5833..5849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5833..5849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5846..5868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5857..5873
                     /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(5881..5911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5926..5947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6064..6081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6235..6823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6315..6334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6994..7854)
                     /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     7617..7636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7855..7959)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttgcca
      901 ccatggagca gaagctgatc tcagaggagg acctggaatt catgaaggga gacaccaggc
      961 atctcaatgg agaggaggac gccggcggga gggaagactc gatcctcgtc aacggggcct
     1021 gcagcgacca gtcctccgac tcgcccccaa tcctggaggc tatccgcacc ccggagatca
     1081 gaggccgaag atcaagctcg cgactctcca agagggaggt gtccagtctg ctaagctaca
     1141 cacaggactt gacaggcgat ggcgacgggg aagatgggga tggctctgac accccagtca
     1201 tgccaaagct cttccgggaa accaggactc gttcagaaag cccagctgtc cgaactcgaa
     1261 ataacaacag tgtctccagc cgggagaggc acaggccttc cccacgttcc acccgaggcc
     1321 ggcagggccg caaccatgtg gacgagtccc ccgtggagtt cccggctacc aggtccctga
     1381 gacggcgggc aacagcatcg gcaggaacgc catggccgtc ccctcccagc tcttacctta
     1441 ccatcgacct cacagacgac acagaggaca cacatgggac gccccagagc agcagtaccc
     1501 cctacgcccg cctagcccag gacagccagc aggggggcat ggagtccccg caggtggagg
     1561 cagacagtgg agatggagac agttcagagt atcaggatgg gaaggagttt ggaatagggg
     1621 acctcgtgtg gggaaagatc aagggcttct cctggtggcc cgccatggtg gtgtcttgga
     1681 aggccacctc caagcgacag gctatgtctg gcatgcggtg ggtccagtgg tttggcgatg
     1741 gcaagttctc cgaggtctct gcagacaaac tggtggcact ggggctgttc agccagcact
     1801 ttaatttggc caccttcaat aagctcgtct cctatcgaaa agccatgtac catgctctgg
     1861 agaaagctag ggtgcgagct ggcaagacct tccccagcag ccctggagac tcattggagg
     1921 accagctgaa gcccatgttg gagtgggccc acgggggctt caagcccact gggatcgagg
     1981 gcctcaaacc caacaacacg caaccagtgg ttaataagtc gaaggtgcgt cgtgcaggca
     2041 gtaggaaatt agaatcaagg aaatacgaga acaagactcg aagacgcaca gctgacgact
     2101 cagccacctc tgactactgc cccgcaccca agcgcctcaa gacaaattgc tataacaacg
     2161 gcaaagaccg aggggatgaa gatcagagcc gagaacaaat ggcttcagat gttgccaaca
     2221 acaagagcag cctggaagat ggctgtttgt cttgtggcag gaaaaacccc gtgtccttcc
     2281 accctctctt tgaggggggg ctctgtcaga catgccggga tcgcttcctt gagctgtttt
     2341 acatgtatga tgacgatggc tatcagtctt actgcactgt gtgctgcgag ggccgagagc
     2401 tgctgctttg cagcaacacg agctgctgcc ggtgtttctg tgtggagtgc ctggaggtgc
     2461 tggtgggcac aggcacagcg gccgaggcca agcttcagga gccctggagc tgttacatgt
     2521 gtctcccgca gcgctgtcat ggcgtcctgc ggcgccggaa ggactggaac gtgcgcctgc
     2581 aggccttctt caccagtgac acggggcttg aatatgaagc ccccaagctg taccctgcca
     2641 ttcccgcagc ccgaaggcgg cccattcgag tcctgtcatt gtttgatggc atcgcgacag
     2701 gctacctagt cctcaaagag ttgggcataa aggtaggaaa gtacgtcgct tctgaagtgt
     2761 gtgaggagtc cattgctgtt ggaaccgtga agcacgaggg gaatatcaaa tacgtgaacg
     2821 acgtgaggaa catcacaaag aaaaatattg aagaatgggg cccatttgac ttggtgattg
     2881 gcggaagccc atgcaacgat ctctcaaatg tgaatccagc caggaaaggc ctgtatgagg
     2941 gtacaggccg gctcttcttc gaattttacc acctgctgaa ttactcacgc cccaaggagg
     3001 gtgatgaccg gccgttcttc tggatgtttg agaatgttgt agccatgaag gttggcgaca
     3061 agagggacat ctcacggttc ctggagtgta atccagtgat gattgatgcc atcaaagttt
     3121 ctgctgctca cagggcccga tacttctggg gcaacctacc cgggatgaac aggcccgtga
     3181 tagcatcaaa gaatgataaa ctcgagctgc aggactgctt ggaatacaat aggatagcca
     3241 agttaaagaa agtacagaca ataaccacca agtcgaactc gatcaaacag gggaaaaacc
     3301 aacttttccc tgttgtcatg aatggcaaag aagatgtttt gtggtgcact gagctcgaaa
     3361 ggatctttgg ctttcctgtg cactacacag acgtgtccaa catgggccgt ggtgcccgcc
     3421 agaagctgct gggaaggtcc tggagcgtgc ctgtcatccg acacctcttc gcccctctga
     3481 aggactactt tgcatgtgaa taggatccgc ggccgctcga gcatgcatct agagggccct
     3541 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     3601 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3661 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3721 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3781 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3841 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3901 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3961 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4021 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4081 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4141 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4201 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4261 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4381 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4441 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4501 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4561 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4621 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4681 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4741 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4801 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4861 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4921 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4981 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5041 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5101 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5161 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5221 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5281 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5341 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5401 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5461 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5521 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5581 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5641 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5761 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5821 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5881 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5941 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6001 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6061 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6661 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7981 acctgacg
//
