LOCUS       Exported                6645 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 6645)
  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..6645
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3346
                     /organism="Homo sapiens"
                     /chromosome="12"
                     /map="12p13.33"
                     /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..1508
                     /label=B41F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3346
                     /codon_start=1
                     /gene="KDM5A"
                     /gene_synonym="RBBP-2; RBBP2; RBP2"
                     /product="lysine-specific demethylase 5A"
                     /EC_number="1.14.11.67"
                     /label=KDM5A
                     /note="retinoblastoma binding protein 2; Jumonji, AT rich 
                     interactive domain 1A (RBP2-like); lysine (K)-specific 
                     demethylase 5A; histone demethylase JARID1A; jumonji/ARID 
                     domain-containing protein 1A; [histone 
                     H3]-trimethyl-L-lysine(4) demethylase 5A"
                     /db_xref="GI:110618244"
                     /db_xref="CCDS:CCDS41736.1"
                     /db_xref="GeneID:5927"
                     /db_xref="HGNC:HGNC:9886"
                     /db_xref="MIM:180202"
                     /protein_id="NP_001036068.1"
                     /translation="VLSPRTDIGVYGSGKNRRKKVKELIEKEKEKDLDLEPLSDLEEGL
                     EETRDTAMVVAVFKEREQKEIEAMHSLRAANLAKMTMVDRIEEVKFCICRKTASGFMLQ
                     CELCKDWFHNSCVPLPKSSSQKKGSSWQAKEVKFLCPLCMRSRRPRLETILSLLVSLQK
                     LPVRLPEGEALQCLTERAMSWQDRARQALATDELSSALAKLSVLSQRMVEQAAREKTEK
                     IISAELQKAAANPDLQGHLPSFQQSAFNRVVSSVSSSPRQTMDYDDEETDSDEDIRETY
                     GYDMKDTASVKSSSSLEPNLFCDEEIPIKSEEVVTHMWTAPSFCAEHAYSSASKSCSQG
                     SSTPRKQPRKSPLVPRSLEPPVLELSPGAKAQLEELMMVGDLLEVSLDETQHIWRILQA
                     THPPSEDRFLHIMEDDSMEEKPLKVKGKDSSEKKRKRKLEKVEQLFGEGKQKSKELKKM
                     DKPRKKKLKLGADKSKELNKLAKKLAKEEERKKKKEKAAAAKVELVKESTEKKREKKVL
                     DIPSKYDWSGAEESDDENAVCAAQNCQRPCKDKVDWVQCDGGCDEWFHQVCVGVSPEMA
                     ENEDYICINCAKKQGPVSPGPAPPPSFIMSYKLPMEDLKETS"
     misc_feature    1493..1494
                     /label=Feature 58
     misc_feature    1607..1609
                     /gene="KDM5A"
                     /gene_synonym="RBBP-2; RBBP2; RBP2"
                     /note="Phosphoserine.
                     /evidence=ECO:0007744|PubMed:18669648, 
                     ECO:0007744|PubMed:19690332, ECO:0007744|PubMed:20068231, 
                     ECO:0007744|PubMed:24275569; propagated from 
                     UniProtKB/Swiss-Prot (P29375.3); phosphorylation site"
     primer_bind     2117..2134
                     /label=B41F-SF1
     primer_bind     complement(3325..3369)
                     /label=nB41F-BsrGI-R
     primer_bind     complement(3325..3367)
                     /label=B41F-BsrGI-R
     misc_feature    3347..3352
                     /label=BsrGI
     CDS             3353..3418
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3358..3458)
                     /label=296
     terminator      3434..3865
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3434..3459
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3513..3536)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3908..4048
                     /label=R-ITR
     rep_origin      4051..4506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4207)
                     /label=F-Seq-Cre-3F1
     promoter        4788..4892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4893..5753
                     /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      5924..6512
                     /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 aggtgctgag
     1501 cccccggacc gacattggtg tatatgggag tggcaaaaat aggaggaaaa aagtaaaaga
     1561 actaatagaa aaagaaaaag aaaaggatct ggacctggag cctctgagtg atctggagga
     1621 aggattggag gaaaccagag atacagccat ggtggtggca gttttcaaag aacgggagca
     1681 aaaagagatt gaagccatgc attctctcag agcagccaac ctagccaaga tgacaatggt
     1741 ggaccgcata gaagaagtaa aattttgcat ttgccgcaag acagccagtg ggtttatgct
     1801 acagtgtgag ctctgcaaag actggttcca taacagctgt gttcctcttc ctaaatcaag
     1861 ttcccaaaaa aaaggatcca gctggcaagc taaagaagta aaattccttt gccctctttg
     1921 tatgcggtct cgaaggccca ggctagagac tattctgtca ctcctggtat cccttcagaa
     1981 gttgcccgta cggttgcctg aaggagaggc cctgcagtgt ttgacagaac gtgctatgag
     2041 ttggcaagat agagcgcggc aggctctagc cacagatgaa ctatcctctg ccctggccaa
     2101 actatctgtg ttgagccagc gtatggtgga acaggcggct cgagaaaaaa ctgaaaagat
     2161 catcagtgca gaactccaaa aagcagctgc caatccagac ttacagggac acttacctag
     2221 tttccagcag tctgctttta accgggtggt gagcagtgtg tcatcttctc ctcgacaaac
     2281 aatggactat gatgatgaag aaacagactc tgatgaagac attcgagaga catatggcta
     2341 cgacatgaag gacacagcca gtgtgaagtc ctctagtagt cttgaaccca atcttttttg
     2401 tgatgaagag attcccatca aatccgagga ggtggtgacc cacatgtgga cagcaccttc
     2461 attttgtgca gagcatgctt attcttctgc ttctaagagt tgttctcaag gttctagcac
     2521 cccaaggaaa caacctcgga agagcccttt ggtgccccga agtttggaac ctccagtgct
     2581 ggagttgtca cctggagcta aggcacaact ggaagaactt atgatggttg gagatctcct
     2641 ggaagtatct ctggacgaga ctcaacacat atggcggatt ttgcaggcca cacacccacc
     2701 ctctgaagac agattcttgc atatcatgga ggatgacagc atggaagaga aaccactaaa
     2761 agtgaaagga aaggactctt cagagaagaa acggaaacgg aagctagaaa aggtagagca
     2821 actttttgga gaaggaaaac agaagtccaa ggagttaaag aaaatggaca aacctagaaa
     2881 gaagaaatta aaattaggtg cagacaaatc aaaggagctg aataaactgg ccaagaaact
     2941 agcaaaagaa gaagagagaa agaaaaagaa ggagaaggct gctgcagcca aagttgaact
     3001 tgtgaaagag agcactgaaa agaaaagaga gaaaaaggtg ctggacatcc cctcaaagta
     3061 tgactggtca ggagcagagg agtctgatga tgagaatgct gtgtgcgcag cacagaactg
     3121 ccaaaggccc tgcaaggaca aggtagactg ggtacaatgt gatggtggct gtgatgagtg
     3181 gtttcatcaa gtttgtgtgg gtgtatctcc agaaatggct gaaaatgaag attacatctg
     3241 tataaactgt gcaaagaagc aggggccagt tagcccaggt ccagcaccac ctccttcctt
     3301 cataatgagc tacaaactac caatggagga tcttaaagag accagttgta cagactacaa
     3361 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     3421 agaattctga tgactcgaga taatcaacct ctggattaca aaatttgtga aagattgact
     3481 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3541 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     3601 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3661 ctgttgggca ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta
     3721 tttgtaacca tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3781 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3841 gggggagatg tgggaggttt tttaaagatc tggagaccgg tgtggcacgt gcggaccgag
     3901 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3961 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4021 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4081 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4141 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     4201 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4261 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4321 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4381 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4441 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4501 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4561 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4621 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4681 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4741 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4801 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4921 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4981 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5041 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5101 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5161 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5221 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5281 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5341 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5401 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5461 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5521 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5701 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
