LOCUS       Exported                6715 bp ds-DNA     circular SYN 16-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6715)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 20, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6715
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3416
                     /organism="Homo sapiens"
                     /chromosome="17"
                     /map="17p13.1"
                     /mol_type="mRNA"
                     /db_xref="taxon:9606"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     primer_bind     141..190
                     /label=F-D1
     primer_bind     147..194
                     /label=F-Mlu-D2
     primer_bind     147..161
                     /label=3'-pcalm-ampF
     primer_bind     150..168
                     /label=P-Calm1-F
     primer_bind     151..200
                     /label=F-D2
     misc_feature    156..276
                     /label=P Calm1
     misc_feature    156..165
                     /label=D1
     primer_bind     161..210
                     /label=F-D3
     primer_bind     complement(161..185)
                     /label=R-D4
     misc_feature    166..175
                     /label=D2
     primer_bind     171..220
                     /label=F-D4
     primer_bind     complement(171..195)
                     /label=R-D5
     misc_feature    176..185
                     /label=D3
     primer_bind     181..230
                     /label=F-D5
     primer_bind     complement(181..205)
                     /label=R-D6
     misc_feature    186..195
                     /label=D4
     primer_bind     191..240
                     /label=F-D6
     primer_bind     complement(191..215)
                     /label=r-d7
     misc_feature    196..205
                     /label=D5
     primer_bind     200..250
                     /label=F-D7
     primer_bind     complement(201..225)
                     /label=R-D8
     misc_feature    206..215
                     /label=D6
     primer_bind     211..260
                     /label=F-D8
     primer_bind     complement(215..236)
                     /label=R-D9
     misc_feature    216..225
                     /label=D7
     primer_bind     222..271
                     /label=F-D9
     primer_bind     complement(222..246)
                     /label=R-D10
     misc_feature    226..235
                     /label=D8
     primer_bind     complement(232..256)
                     /label=R-D11
     misc_feature    236
                     /label=+1
     misc_feature    237..246
                     /label=D9
     primer_bind     complement(242..266)
                     /label=R-D12
     misc_feature    247..256
                     /label=D10
     misc_feature    257..266
                     /label=D11
     primer_bind     complement(260..287)
                     /label=2B-1R
     primer_bind     complement(261..276)
                     /label=P-Calm1-R
     misc_feature    267..276
                     /label=D12
     primer_bind     271..296
                     /label=2B-2F
     primer_bind     complement(277..298)
                     /label=Primer 1
     misc_feature    277..282
                     /label=BsiWI
     primer_bind     283..302
                     /label=Primer 5
     CDS             288..1318
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    548
                     /label=silent mut
     primer_bind     complement(616..650)
                     /label=R-Cre-R119STOP
     primer_bind     complement(632..651)
                     /label=R-CRE-L115-BamHI
     primer_bind     636..670
                     /label=F-Cre-R119STOP
     primer_bind     1093..1116
                     /label=F-Seq-Cre-3F1
     misc_feature    1093..1098
                     /label=ClaI
     misc_feature    1224
                     /label=mut
     primer_bind     1262..1279
                     /label=cre-SF
     primer_bind     complement(1296..1318)
                     /label=R-T5-Cre
     intron          1320..1360
                     /label=Splicing donor
                     /note="Splicing donor"
     primer_bind     complement(1338..1367)
                     /label=2B-2R
     primer_bind     complement(1341..1385)
                     /label=Primer 7
     primer_bind     complement(1341..1385)
                     /label=Primer 6
     primer_bind     1350..1387
                     /label=2B-3F
     primer_bind     complement(1356..1380)
                     /label=Primer 3
     primer_bind     1367..1401
                     /label=Primer 8
     primer_bind     1367..1401
                     /label=Primer 2
     primer_bind     1367..1401
                     /label=F-T5-JT+2272
     protein_bind    1367..1400
                     /label=loxJTZ17?2272?
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1380..1387
                     /label=2272
     intron          1401..1492
                     /label=Splicing acceptor
                     /note="Splicing acceptor"
     primer_bind     1401..1420
                     /label=Primer 4
     primer_bind     1459..1478
                     /label=T0060_PIRES-F
     primer_bind     complement(1468..1492)
                     /label=#57-R-SA-3R1
     primer_bind     1477..1510
                     /label=A23F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3416
                     /codon_start=1
                     /gene="KDM6B"
                     /gene_synonym="JMJD3; NEDCFSA"
                     /product="lysine-specific demethylase 6B isoform 2"
                     /EC_number="1.14.11.68"
                     /label=KDM6B
                     /note="isoform 2 is encoded by transcript variant 2;
                     jumonji domain containing 3, histone lysine demethylase; 
                     lysine (K)-specific demethylase 6B; lysine-specific 
                     demethylase 6B; jmjC domain-containing protein 3; jumonji 
                     domain-containing protein 3; [histone 
                     H3]-trimethyl-L-lysine(27) demethylase 6B"
                     /db_xref="GI:1145994701"
                     /db_xref="CCDS:CCDS86569.1"
                     /db_xref="GeneID:23135"
                     /db_xref="HGNC:HGNC:29012"
                     /db_xref="MIM:611577"
                     /protein_id="NP_001335645.1"
                     /translation="AKAKAKVPKEKSRRVLGNLDLQSEEIQGREKSRPDLGGASKAKPP
                     TAPAPPSAPAPSAQPTPPSASVPGKKAREEAPGPPGVSRADMLKLRSLSEGPPKELKIR
                     LIKVESGDKETFIASEVEERRLRMADLTISHCAADVVRASRNAKVKGKFRESYLSPAQS
                     VKPKINTEEKLPREKLNPPTPSIYLESKRDAFSPVLLQFCTDPRNPITVIRGLAGSLRL
                     NLGLFSTKTLVEASGEHTVEVRTQVQQPSDENWDLTGTRQIWPCESSRSHTTIAKYAQY
                     QASSFQESLQEEKESEDEESEEPDSTTGTPPSSAPDPKNHHIIKFGTNIDLSDAKRWKP
                     QLQELLKLPAFMRVTSTGNMLSHVGHTILGMNTVQLYMKVPGSRTPGHQENNNFCSVNI
                     NIGPGDCEWFAVHEHYWETISAFCDRHGVDYLTGSWWPILDDLYASNIPVYRFVQRPGD
                     LVWINAGTVHWVQATGWCNNIAWNVGPLTAYQYQLALERYEWNEVKNVKSIVPMIHVSW
                     NVARTVKISDPDLFKMIKFCLLQSMKHCQVQRESLVRAGKKIAYQGRVKDEPAYYCNEC
                     DVEVFNILFVTSENGSRNTYLVHCEGCARRRSAGLQGVVVLEQYRTEELAQAYDAFTLA
                     PASTSR"
     misc_feature    1493..1494
                     /label=Feature 41
     primer_bind     2048..2064
                     /label=A23F-SF
     primer_bind     2085..2102
                     /label=A23F-BsrGI-1F
     primer_bind     complement(2147..2163)
                     /label=23B-KDM6B-SR2
     primer_bind     complement(2717..2733)
                     /label=23B-KDM6B-SR
     primer_bind     2766..2776
                     /label=P-Calm1-R
     primer_bind     complement(2766..2776)
                     /label=F-D9
     primer_bind     complement(3402..3437)
                     /label=A23F-BsrGI-R
     primer_bind     complement(3403..3440)
                     /label=A23F-BsrGI-1R
     primer_bind     complement(3403..3422)
                     /label=Primer 1
     misc_feature    3417..3422
                     /label=BsrGI
     CDS             3423..3488
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3428..3528)
                     /label=296
     terminator      3504..3935
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3504..3529
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3583..3606)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3978..4118
                     /label=R-ITR
     rep_origin      4121..4576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4268..4277)
                     /label=F-Seq-Cre-3F1
     promoter        4858..4962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4963..5823
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="HSNMYPLMRQ*P**MLQ*Y*KRKSMSIQHFRVALIPFFAAFCLPV
                     FAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLS
                     RIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTT
                     IGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLT
                     LASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIV
                     VIYTTG"
     rep_origin      5994..6582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcgctc tgcgcgaggc ggctccgccg
      181 cggcggaggg atacggcgca ccatatatat atcgcggggc gcagactcgc gctccggcag
      241 tggtgctggg agtgtcgtgg acgccgtgcc gttactcgta cgccaccatg tccaatttac
      301 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      361 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      421 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      481 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      541 taaaaaccat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      601 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      661 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      721 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      781 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      841 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      901 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      961 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
     1021 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1081 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1141 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1201 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgta atgaactata
     1261 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagg
     1321 taagtatcaa ggttacaaga caggtttaag gagaccaata actagtataa cttcgtatag
     1381 gatactttat agcaatttat gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1441 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac agggcaaagg
     1501 ccaaggccaa ggtccccaaa gaaaagagcc gccgggtgct ggggaacctg gacctgcaga
     1561 gcgaggagat ccagggtcgt gagaagtccc ggcccgatct tggcggggcc tccaaggcca
     1621 agccacccac agctccagcc cctccatcag ctcctgcacc ttctgcccag cccacacccc
     1681 cgtcagcctc tgtccctgga aagaaggctc gggaggaagc cccagggcca ccgggtgtca
     1741 gccgggccga catgctgaag ctgcgctcac ttagtgaggg gccccccaag gagctgaaga
     1801 tccggctcat caaggtagag agtggtgaca aggagacctt tatcgcctct gaggtggaag
     1861 agcggcggct gcgcatggca gacctcacca tcagccactg tgctgctgac gtcgtgcgcg
     1921 ccagcaggaa tgccaaggtg aaagggaagt ttcgagagtc ctacctttcc cctgcccagt
     1981 ctgtgaaacc gaagatcaac actgaggaga agctgccccg ggaaaaactc aaccccccta
     2041 cacccagcat ctatctggag agcaaacggg atgccttctc acctgtcctg ctgcagttct
     2101 gtacagaccc tcgaaatccc atcacagtga tccggggcct ggcgggctcc ctgcggctca
     2161 acttgggcct cttctccacc aagaccctgg tggaagcgag tggcgaacac accgtggaag
     2221 ttcgcaccca ggtgcagcag ccctcagatg agaactggga tctgacaggc actcggcaga
     2281 tctggccttg tgagagctcc cgttcccaca ccaccattgc caagtacgca cagtaccagg
     2341 cctcatcctt ccaggagtct ctgcaggagg agaaggagag tgaggatgag gagtcagagg
     2401 agccagacag caccactgga acccctccta gcagcgcacc agacccgaag aaccatcaca
     2461 tcatcaagtt tggcaccaac atcgacttgt ctgatgctaa gcggtggaag ccccagctgc
     2521 aggagctgct aaagctgccc gccttcatgc gggtaacatc cacgggcaac atgctgagcc
     2581 acgtgggcca caccatcctg ggcatgaaca cggtgcagct gtacatgaag gtgcccggca
     2641 gccgaacgcc aggccaccag gagaataaca acttctgctc cgtcaacatc aacattggcc
     2701 caggcgactg cgagtggttc gcggtgcacg agcactactg ggagaccatc agcgctttct
     2761 gtgatcggca cggcgtggac tacttgacgg gttcctggtg gccaatcctg gatgatctct
     2821 atgcatccaa tattcctgtg taccgcttcg tgcagcgacc cggagacctc gtgtggatta
     2881 atgcggggac tgtgcactgg gtgcaggcca ccggctggtg caacaacatt gcctggaacg
     2941 tggggcccct caccgcctat cagtaccagc tggccctgga acgatacgag tggaatgagg
     3001 tgaagaacgt caaatccatc gtgcccatga ttcacgtgtc atggaacgtg gctcgcacgg
     3061 tcaaaatcag cgaccccgac ttgttcaaga tgatcaagtt ctgcctgctg cagtccatga
     3121 agcactgcca ggtgcaacgc gagagcctgg tgcgggcagg gaagaaaatc gcttaccagg
     3181 gccgtgtcaa ggacgagcca gcctactact gcaacgagtg cgatgtggag gtgtttaaca
     3241 tcctgttcgt gacaagtgag aatggcagcc gcaacacgta cctggtacac tgcgagggct
     3301 gtgcccggcg ccgcagcgca ggcctgcagg gcgtggtggt gctggagcag taccgcactg
     3361 aggagctggc tcaggcctac gacgccttca cgctggcccc agccagcacg tcgcgatgta
     3421 cagactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3481 atgacaagta agaattctga tgactcgaga taatcaacct ctggattaca aaatttgtga
     3541 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3601 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3661 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3721 aggggctcgg ctgttgggca ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc
     3781 tattgcttta tttgtaacca tctagcttta tttgtgaaat ttgtgatgct attgctttat
     3841 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3901 ttcaggttca gggggagatg tgggaggttt tttaaagatc tggagaccgg tgtggcacgt
     3961 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4021 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4081 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4141 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4261 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4381 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4501 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     4561 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4621 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
