LOCUS       Exported                5538 bp ds-DNA     circular SYN 29-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAK170_2x3APWWP-IFP2.0C
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5538)
  AUTHORS   Jeltsch Group
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 29, 2024 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5538
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    525..612
                     /label=sequenced
     misc_feature    616..1044
                     /label=PWWP domain
     CDS             616..1044
                     /codon_start=1
                     /product="DNA methyltransferase 3A"
                     /label=DNMT3A
                     /translation="EYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRW
                     VMWFGDGKFSVVCVEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPV
                     CHDSDESDTAKAVEVQNKPMIEWALGGFQPSGPKGLEPP"
     misc_feature    1045..1053
                     /label=18aa C50 linker
                     /label=50aa C50 linker
                     /note="Copied from rg2104_ChePep3_hRas"
     misc_feature    1045..1053
                     /label=sequenced
     CDS             1054..1074
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1071..1080
                     /label=sequenced
     misc_feature    1084..1512
                     /label=PWWP domain
     CDS             1084..1512
                     /codon_start=1
                     /product="DNA methyltransferase 3A"
                     /label=DNMT3A
                     /translation="EYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRW
                     VMWFGDGKFSVVCVEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPV
                     CHDSDESDTAKAVEVQNKPMIEWALGGFQPSGPKGLEPP"
     misc_feature    1513..1566
                     /label=sequenced
     misc_feature    1513..1566
                     /label=18aa C50 linker
                     /label=50aa C50 linker
                     /note="Copied from rg2104_ChePep3_hRas"
     CDS             1566..2133
                     /codon_start=1
                     /product="bacteriophytochrome-based monomeric infrared 
                     fluorescent protein (Yu et al., 2014)"
                     /label=IFP2.0
                     /translation="DSIGPHALRNAMFALESAPNLRALAEVATQTVRELSGFDRVMLYK
                     FAPDATGEVIAEARREGMQAYLGHRFPASTTPAQARALYTRHLLRLTADTRAAAVPLDP
                     VLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGGQLWGLIVCHHQTPY
                     VLPPDLRTTLEYLGRLLSLQVQRKEA"
     misc_feature    1566
                     /label=N132
     misc_feature    1567..2133
                     /label=C133
     misc_feature    2134..2136
                     /label=Synthetic stop
     CDS             2134..2136
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation=""
     misc_feature    2137..2202
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    2211..2221
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     polyA_signal    2326..2447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2454..2909)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2936..3040
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3042..3399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3250..3385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3434..4228
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4460..4507
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    4460..4507
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      4836..5424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggagta tgaggatggc cggggctttg gcattgggga gctggtgtgg
      661 gggaaactgc ggggcttctc ctggtggcca ggccgcattg tgtcttggtg gatgacgggc
      721 cggagccgag cagctgaagg cacccgctgg gtcatgtggt tcggagacgg caaattctca
      781 gtggtgtgtg ttgagaagct gatgccgctg agctcgtttt gcagtgcgtt ccaccaggcc
      841 acgtacaaca agcagcccat gtaccgcaaa gccatctacg aggtcctgca ggtggccagc
      901 agccgcgcgg ggaagctgtt cccggtgtgc cacgacagcg atgagagtga cactgccaag
      961 gccgtggagg tgcagaacaa gcccatgatt gaatgggccc tggggggctt ccagccttct
     1021 ggccctaagg gcctggagcc accaactgga tcgcccaaga aaaaacgcaa ggtggccacc
     1081 atggagtatg aggatggccg gggctttggc attggggagc tggtgtgggg gaaactgcgg
     1141 ggcttctcct ggtggccagg ccgcattgtg tcttggtgga tgacgggccg gagccgagca
     1201 gctgaaggca cccgctgggt catgtggttc ggagacggca aattctcagt ggtgtgtgtt
     1261 gagaagctga tgccgctgag ctcgttttgc agtgcgttcc accaggccac gtacaacaag
     1321 cagcccatgt accgcaaagc catctacgag gtcctgcagg tggccagcag ccgcgcgggg
     1381 aagctgttcc cggtgtgcca cgacagcgat gagagtgaca ctgccaaggc cgtggaggtg
     1441 cagaacaagc ccatgattga atgggccctg gggggcttcc agccttctgg ccctaagggc
     1501 ctggagccac caactggatc gtcacaaacc tcgggttcat ccacgtcgga aggccaggag
     1561 ggcacggact ctattggacc ccacgctctg agaaatgcaa tgttcgctct ggagagtgct
     1621 cctaatctcc gggcactggc tgaagtcgca acccaaacag tccgggaact gtcaggtttc
     1681 gaccgggtga tgctgtacaa gtttgcacca gacgcaacag gagaggttat tgccgaagca
     1741 aggcgcgagg gcatgcaggc ttacctcggg cataggtttc ccgcatccac cacccctgca
     1801 caagctaggg ccctctacac aagacacctg ctccggctga ccgcagacac cagggctgca
     1861 gcagtgcccc tcgaccccgt gctgaatccc cagacaaatg ctcctacacc tctgggcgga
     1921 gctgtcctca gagctacatc cccaatgcac atgcagtacc tgaggaatat gggagtgggc
     1981 tcctccctga gcgtcagcgt cgtggtcggc ggccagctgt ggggactgat tgtctgccac
     2041 catcagacac cctacgtgct gccacccgat ctgcggacca ccctggagta tctggggagg
     2101 ctcctgtccc tgcaggtgca gaggaaagaa gcctaatccg gactcagatc tcgagctcaa
     2161 gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     2221 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2281 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2341 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2401 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2461 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2521 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2581 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2641 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2701 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2761 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2821 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2881 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2941 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3001 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3301 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3601 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3961 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4381 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4441 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4501 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4561 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4621 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4981 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5521 cgtattaccg ccatgcat
//
