LOCUS       Exported                6906 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 6906)
  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..6906
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3619
                     /organism="Homo sapiens"
                     /chromosome="X"
                     /map="Xp11.3"
                     /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..1509
                     /label=B13F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3619
                     /codon_start=1
                     /gene="KDM6A"
                     /gene_synonym="bA386N14.2; KABUK2; UTX"
                     /product="lysine-specific demethylase 6A isoform 1"
                     /EC_number="1.14.11.68"
                     /label=KDM6A
                     /note="isoform 1 is encoded by transcript variant 1; 
                     ubiquitously-transcribed TPR gene on the X chromosome; 
                     ubiquitously transcribed tetratricopeptide repeat protein 
                     X-linked; lysine (K)-specific demethylase 6A; bA386N14.2 
                     (ubiquitously transcribed X chromosome tetratricopeptide 
                     repeat protein (UTX)); histone demethylase UTX; [histone 
                     H3]-trimethyl-L-lysine(27) demethylase 6A"
                     /db_xref="GI:611434998"
                     /db_xref="CCDS:CCDS94599.1"
                     /db_xref="GeneID:7403"
                     /db_xref="HGNC:HGNC:12637"
                     /db_xref="MIM:300128"
                     /protein_id="NP_001278344.1"
                     /translation="GAALNHLSSHTATSGGQQGITLTKESKPSGNILTVPETSRHTGET
                     PNSTASVEGLPNHVHQMTADAVCSPSHGDSKSPGLLSSDNPQLSALLMGKANNNVGTGT
                     CDKVNNIHPAVHTKTDNSVASSPSSAISTATPSPKSTEQTTTNSVTSLNSPHSGLHTIN
                     GEGMEESQSPMKTDLLLVNHKPSPQIIPSMSVSIYPSSAEVLKACRNLGKNGLSNSSIL
                     LDKCPPPRPPSSPYPPLPKDKLNPPTPSIYLENKRDAFFPPLHQFCTNPNNPVTVIRGL
                     AGALKLDLGLFSTKTLVEANNEHMVEVRTQLLQPADENWDPTGTKKIWHCESNRSHTTI
                     AKYAQYQASSFQESLREENEKRSHHKDHSDSESTSSDNSGRRRKGPFKTIKFGTNIDLS
                     DDKKWKLQLHELTKLPAFVRVVSAGNLLSHVGHTILGMNTVQLYMKVPGSRTPGHQENN
                     NFCSVNINIGPGDCEWFVVPEGYWGVLNDFCEKNNLNFLMGSWWPNLEDLYEANVPVYR
                     FIQRPGDLVWINAGTVHWVQAIGWCNNIAWNVGPLTACQYKLAVERYEWNKLQSVKSIV
                     PMVHLSWNMARNIKVSDPKLFEMIKYCLLRTLKQCQTLREALIAAGKEIIWHGRTKEEP
                     AHYCSICEVEVFDLLFVTNESNSRKTYIVHCQDCARKTSGNLENFVVLEQYKMEDLMQV
                     YDQFTLAPPLPSASS"
     misc_feature    1493..1494
                     /label=Feature 43
     primer_bind     complement(3600..3642)
                     /label=nB13F-BsrGI-R
     primer_bind     complement(3600..3640)
                     /label=B13F-BsrGI-R
     misc_feature    3620..3625
                     /label=BsrGI
     CDS             3626..3691
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3631..3719)
                     /label=296
     terminator      3695..4126
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3695..3720
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3774..3797)
                     /label=R-Seq-W3SL-5R1
     misc_feature    4169..4309
                     /label=R-ITR
     rep_origin      4312..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4459..4468)
                     /label=F-Seq-Cre-3F1
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5154..6014
                     /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      6185..6773
                     /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 agggggctgc
     1501 tctcaatcac ctctcctctc acactgctac ctcaggtgga caacaaggca ttaccttaac
     1561 caaagagagc aagccttcag gaaacatatt gacggtgcct gaaacaagca ggcacactgg
     1621 agagacacct aacagcactg ccagtgtcga gggacttcct aatcatgtcc atcagatgac
     1681 ggcagatgct gtttgcagtc ctagccatgg agattctaag tcaccaggtt tactaagttc
     1741 agacaatcct cagctctctg ccttgttgat gggaaaagcc aataacaatg tgggtactgg
     1801 aacctgtgac aaagtcaata acatccaccc agctgttcat acaaagactg ataactctgt
     1861 tgcctcttca ccatcttcag ccatttcaac agcaacacct tctccaaaat ccactgagca
     1921 gacaaccaca aacagtgtta ccagccttaa cagccctcac agtgggctac acacaattaa
     1981 tggagaaggg atggaagaat ctcagagccc catgaaaaca gatctgcttc tggttaacca
     2041 caaacctagt ccacagatca taccatcaat gtctgtgtcc atatacccca gctcagcaga
     2101 agttctgaag gcatgcagga atctaggtaa aaatggctta tctaacagta gcattttgtt
     2161 ggataaatgt ccacctccaa gaccaccatc ttcaccatac cctcccttgc caaaggacaa
     2221 gttgaatcca cctacaccta gtatttactt ggaaaataaa cgtgatgctt tctttcctcc
     2281 attacatcaa ttttgtacaa atccgaacaa ccctgttaca gtaatacgtg gccttgctgg
     2341 agctcttaag ttagacctgg gacttttctc tactaaaact ttggtggaag ctaacaatga
     2401 acatatggta gaagtgagga cacagttgtt gcagccagca gatgaaaact gggatcccac
     2461 tggaacaaag aaaatctggc attgtgaaag taatagatct catactacaa ttgctaaata
     2521 tgcacagtac caggcctcct cattccagga atcattgaga gaagaaaatg aaaaaagaag
     2581 tcatcataaa gaccactcag atagtgaatc tacatcgtca gataattctg ggaggaggag
     2641 gaaaggaccc tttaaaacca taaagtttgg gaccaatatt gacctatctg atgacaaaaa
     2701 gtggaagttg cagctacatg agctgactaa acttcctgct tttgtgcgtg tcgtatcagc
     2761 aggaaatctt ctaagccatg ttggtcatac catattgggc atgaacacag ttcaactata
     2821 catgaaagtt ccagggagca gaacaccagg tcatcaggaa aataacaact tctgttcagt
     2881 taacataaat attggcccag gtgactgtga atggtttgtt gttcctgaag gttactgggg
     2941 tgttctgaat gacttctgtg aaaaaaataa tttgaatttc ctaatgggtt cttggtggcc
     3001 caatcttgaa gatctttatg aagcaaatgt tccagtgtat aggtttattc agcgacctgg
     3061 agatttggtc tggataaatg caggcactgt tcattgggtt caggctattg gctggtgcaa
     3121 caacattgct tggaatgttg gtccacttac agcctgccag tataaattgg cagtggaacg
     3181 gtacgaatgg aacaaattgc aaagtgtgaa gtcaatagta cccatggttc atctttcctg
     3241 gaatatggca cgaaatatca aggtctcaga tccaaagctt tttgaaatga ttaagtattg
     3301 tcttctaaga actctgaagc aatgtcagac attgagggaa gctctcattg ctgcaggaaa
     3361 agagattata tggcatgggc ggacaaaaga agaaccagct cattactgta gcatttgtga
     3421 agtggaggtt tttgatctgc tttttgtcac taatgagagt aattcacgaa agacctacat
     3481 agtacattgc caagattgtg cacgaaaaac aagcggaaac ttggaaaact ttgtggtgct
     3541 agaacagtac aaaatggagg acctgatgca agtctatgac caatttacat tagctcctcc
     3601 attaccatcc gcctcatctt gtacagacta caaagaccat gacggtgatt ataaagatca
     3661 tgacatcgac tacaaggatg acgatgacaa gtaactcgag ataatcaacc tctggattac
     3721 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3781 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3841 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     3901 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa
     3961 atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa tttgtgatgc
     4021 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4081 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagat ctggagaccg
     4141 gtgtggcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4201 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4261 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4321 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4381 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4441 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4501 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4561 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4621 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4681 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4741 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4801 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4861 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4921 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4981 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5041 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5461 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5761 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6901 acatgt
//
