LOCUS       sequence-133144-        7497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7497)
  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 7497)
  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: "J Cell 
            Biochem."; date: "2005-08-1"; volume: "95(5)"; pages: "902-17"
COMMENT     Sequence Label: pcDNA3/Myc-DNMT3A2
FEATURES             Location/Qualifiers
     source          1..7497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2832..2841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2841..2870
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4011..4943
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     promoter        complement(4987..5005)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4988..5005)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5025..5042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5031..5255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5301..5729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5388..5407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5598..5619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5738..5758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5743..6072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5923..6058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5985..6004
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6139..6933
                     /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(6193..6212)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6803..6822
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7107..7228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7144..7163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7198..7217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7277..7293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7277..7293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7290..7312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7301..7317
                     /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(7325..7331,7332..7349,7350..7355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7370..7391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct tgccaccatg gagcagaagc tgatctcaga ggaggacctg gaattcatga
     2881 atgctgtgga agaaaaccag gggcccgggg agtctcagaa ggtggaggag gccagccctc
     2941 ctgctgtgca gcagcccact gaccccgcat cccccactgt ggctaccacg cctgagcccg
     3001 tggggtccga tgctggggac aagaatgcca ccaaagcagg cgatgacgag ccagagtacg
     3061 aggacggccg gggctttggc attggggagc tggtgtgggg gaaactgcgg ggcttctcct
     3121 ggtggccagg ccgcattgtg tcttggtgga tgacgggccg gagccgagca gctgaaggca
     3181 cccgctgggt catgtggttc ggagacggca aattctcagt ggtgtgtgtt gagaagctga
     3241 tgccgctgag ctcgttttgc agtgcgttcc accaggccac gtacaacaag cagcccatgt
     3301 accgcaaagc catctacgag gtcctgcagg tggccagcag ccgcgcgggg aagctgttcc
     3361 cggtgtgcca cgacagcgat gagagtgaca ctgccaaggc cgtggaggtg cagaacaagc
     3421 ccatgattga atgggccctg gggggcttcc agccttctgg ccctaagggc ctggagccac
     3481 cagaagaaga gaagaatccc tacaaagaag tgtacacgga catgtgggtg gaacctgagg
     3541 cagctgccta cgcaccacct ccaccagcca aaaagccccg gaagagcaca gcggagaagc
     3601 ccaaggtcaa ggagattatt gatgagcgca caagagagcg gctggtgtac gaggtgcggc
     3661 agaagtgccg gaacattgag gacatctgca tctcctgtgg gagcctcaat gttaccctgg
     3721 aacaccccct cttcgttgga ggaatgtgcc aaaactgcaa gaactgcttt ctggagtgtg
     3781 cgtaccagta cgacgacgac ggctaccagt cctactgcac catctgctgt gggggccgtg
     3841 aggtgctcat gtgcggaaac aacaactgct gcaggtgctt ttgcgtggag tgtgtggacc
     3901 tcttggtggg gccgggggct gcccaggcag ccattaagga agacccctgg aactgctaca
     3961 tgtgcgggca caagggtacc tacgggctgc tgcggcggcg agaggactgg ccctcccggc
     4021 tccagatgtt cttcgctaat aaccacgacc aggaatttga ccctccaaag gtttacccac
     4081 ctgtcccagc tgagaagagg aagcccatcc gggtgctgtc tctctttgat ggaatcgcta
     4141 cagggctcct ggtgctgaag gacttgggca ttcaggtgga ccgctacatt gcctcggagg
     4201 tgtgtgagga ctccatcacg gtgggcatgg tgcggcacca ggggaagatc atgtacgtcg
     4261 gggacgtccg cagcgtcaca cagaagcata tccaggagtg gggcccattc gatctggtga
     4321 ttgggggcag tccctgcaat gacctctcca tcgtcaaccc tgctcgcaag ggcctctacg
     4381 agggcactgg ccggctcttc tttgagttct accgcctcct gcatgatgcg cggcccaagg
     4441 agggagatga tcgccccttc ttctggctct ttgagaatgt ggtggccatg ggcgttagtg
     4501 acaagaggga catctcgcga tttctcgagt ccaaccctgt gatgattgat gccaaagaag
     4561 tgtcagctgc acacagggcc cgctacttct ggggtaacct tcccggtatg aacaggccgt
     4621 tggcatccac tgtgaatgat aagctggagc tgcaggagtg tctggagcat ggcaggatag
     4681 ccaagttcag caaagtgagg accattacta cgaggtcaaa ctccataaag cagggcaaag
     4741 accagcattt tcctgtcttc atgaatgaga aagaggacat cttatggtgc actgaaatgg
     4801 aaagggtatt tggtttccca gtccactata ctgacgtctc caacatgagc cgcttggcga
     4861 ggcagagact gctgggccgg tcatggagcg tgccagtcat ccgccacctc ttcgctccgc
     4921 tgaaggagta ttttgcgtgt gtgtaaggat ccgcggccgc tcgagcatgc atctagaggg
     4981 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     5041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5821 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6001 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6061 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6121 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6181 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6241 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6301 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6361 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6421 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6481 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6541 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6601 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6661 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6721 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6781 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6841 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6901 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6961 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7021 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7081 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     7141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7201 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7261 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7321 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7381 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7441 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
