LOCUS       Exported                5959 bp ds-DNA     circular SYN 02-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5959)
  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..5959
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..2672
                     /organism="Homo sapiens"
                     /chromosome="1"
                     /map="1q32.1"
                     /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..1508
                     /label=A15F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..2672
                     /codon_start=1
                     /gene="KDM5B"
                     /gene_synonym="CT31; JARID1B; MRT65; PLU-1; PLU1; PPP1R98;
                     PUT1; RBBP2H1A; RBP2-H1"
                     /product="lysine-specific demethylase 5B isoform 2"
                     /EC_number="1.14.11.67"
                     /label=KDM5B
                     /note="isoform 2 is encoded by transcript variant 2;
                     putative DNA/chromatin binding motif; cancer/testis antigen
                     31; jumonji, AT rich interactive domain 1B; 
                     retinoblastoma-binding protein 2, homolog 1A; histone 
                     demethylase JARID1B; jumonji/ARID domain-containing protein
                     1B; retinoblastoma-binding protein 2 homolog 1; protein 
                     phosphatase 1, regulatory subunit 98; lysine (K)-specific 
                     demethylase 5B; [histone H3]-trimethyl-L-lysine(4) 
                     demethylase 5B"
                     /db_xref="GI:57242796"
                     /db_xref="CCDS:CCDS30974.1"
                     /db_xref="GeneID:10765"
                     /db_xref="HGNC:HGNC:18039"
                     /db_xref="MIM:605393"
                     /protein_id="NP_006609.3"
                     /translation="EMEALQSLRLANEGKLLSPLQDVDIKICLCQKAPAAPMIQCELCR
                     DAFHTSCVAVPSISQGLRIWLCPHCRRSEKPPLEKILPLLASLQRIRVRLPEGDALRYM
                     IERTVNWQHRAQQLLSSGNLKFVQDRVGSGLLYSRWQASAGQVSDTNKVSQPPGTTSFS
                     LPDDWDNRTSYLHSPFSTGRSCIPLHGVSPEVNELLMEAQLLQVSLPEIQELYQTLLAK
                     PSPAQQTDRSSPVRPSSEKNDCCRGKRDGINSLERKLKRRLEREGLSSERWERVKKMRT
                     PKKKKIKLSHPKDMNNFKLERERSYELVRSAETHSLPSDTSYSEQEDSEDEDAICPAVS
                     CLQPEGDEVDWVQCDGSCNQWFHQVCVGVSPEMAEKEDYICVRCTVKDAPSRK"
     misc_feature    1493..1494
                     /label=Feature 47
     primer_bind     complement(2657..2693)
                     /label=A15F-BsrGI-R
     misc_feature    2673..2678
                     /label=BsrGI
     CDS             2679..2744
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2684..2772)
                     /label=296
     terminator      2748..3179
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     2748..2773
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(2827..2850)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3222..3362
                     /label=R-ITR
     rep_origin      3365..3820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3512..3521)
                     /label=F-Seq-Cre-3F1
     promoter        4102..4206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4207..5067
                     /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      5238..5826
                     /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 agggaaatgg
     1501 aagccttgca gtctctcaga ctcgccaatg aagggaaatt gctgtcgcct ctccaagatg
     1561 tggatataaa aatctgccta tgtcagaagg ccccagctgc ccctatgatt caatgtgaac
     1621 tctgcaggga tgctttccac accagttgtg tggcggtacc cagtatttca cagggcctgc
     1681 gaatctggct ttgtccccat tgtcggaggt cagagaaacc tccattagag aaaattctgc
     1741 ccctgctcgc ctcccttcag cgtatccgag ttcgccttcc tgagggagat gcacttcgat
     1801 atatgattga aagaaccgtg aactggcagc acagagccca gcaactgctt tcgtcaggga
     1861 atcttaaatt tgtgcaagat cgagtgggct caggactgtt atatagcaga tggcaagcct
     1921 cagcaggaca ggtgtcagac acaaacaagg tatctcaacc tcctggcaca acatcatttt
     1981 ctttgcctga tgactgggac aacagaacct catatttgca ctcccccttc tcaactggac
     2041 gaagttgtat ccccctccat ggtgttagtc cagaagtgaa tgaactattg atggaagccc
     2101 agctgctcca ggtatccctt cctgaaattc aggaacttta ccagacttta cttgcaaagc
     2161 caagccctgc tcagcagact gaccgaagct caccagtgag acccagcagt gagaagaatg
     2221 actgttgccg agggaagcga gatggaatta acagtcttga gagaaaactg aagagacgcc
     2281 tggaaagaga gggcctctcc agtgagcggt gggaacgagt taagaaaatg cggaccccca
     2341 aaaagaagaa aatcaaactg agccacccca aggacatgaa caatttcaag ttagagagag
     2401 agcgtagcta tgaattagtt cgttctgctg aaactcattc cctgccctca gacacatcct
     2461 attccgaaca ggaagactct gaggatgaag atgccatctg cccagctgtg agctgcctgc
     2521 agccagaagg agatgaggtg gactgggtcc agtgtgatgg cagctgcaat cagtggtttc
     2581 atcaggtctg tgttggtgtc tccccagaga tggcagagaa agaagactac atctgtgtgc
     2641 gctgtactgt gaaggacgca ccaagccgaa agtgtacaga ctacaaagac catgacggtg
     2701 attataaaga tcatgacatc gactacaagg atgacgatga caagtaactc gagataatca
     2761 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2821 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2881 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2941 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3001 gtttatttgt gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg
     3061 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3121 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3181 gatctggaga ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3241 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3301 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3361 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3421 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3481 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3721 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3781 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3841 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3901 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3961 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4021 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4081 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4141 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4201 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4261 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4321 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4381 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4441 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4501 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4561 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4621 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4681 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4741 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4801 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4861 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4921 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4981 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5041 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5101 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5161 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5221 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5461 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5521 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5581 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5641 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5701 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5761 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5821 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5881 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5941 tggccttttg ctcacatgt
//
