LOCUS       sequence-410384-        5074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MDK protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5074)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 5074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: MDKA-c012
FEATURES             Location/Qualifiers
     source          1..5074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          640..1656
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1580..1602
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1664..1683
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1738..1747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2239..2283
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2296..2313
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2425..2444)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2490..2545
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2544..2563)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2906..2922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2906..2922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2919..2941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2930..2946
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2954..2960,2961..2978,2979..2984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2999..3020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3149..3166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3320..3908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3400..3419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4079..4870,4871..4939))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4702..4721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4940..5044)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggggcgcgc gccaggcggg gcggggcgag gcggagaggt
      541 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      601 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      661 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      721 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      781 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      841 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
      901 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      961 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1021 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     1081 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     1141 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     1201 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1261 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1321 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1381 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1441 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1501 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1561 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1621 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1681 gtgctgtctc atcattttgg caaagaattg cggccgtctc aggccgaatt caagcttgcc
     1741 accatgggga tccttcccag ccctgggatg cctgcgctgc tctccctcgt gagccttctc
     1801 tccgtgctgc tgatgggttg cgtagctgaa accggtgccg ccgtcgccaa aaagaaagat
     1861 aaggtgaaga aaggtggtcc tggatcagaa tgtgctgaat gggcatgggg accatgtaca
     1921 ccaagcagca aggattgcgg cgtgggtttc cgcgagggca cctgcggggc ccagacccag
     1981 cgcatccggt gcagggtgcc ctgcaactgg aagaaggagt ttggagccga ctgcaagtac
     2041 aagtttgaga actggggtgc gtgtgatggg ggcacaggca ccaaagtccg ccaaggcacc
     2101 ctgaagaagg cgcgctacaa tgctcagtgc caggagacca tccgcgtcac caagccctgc
     2161 acccccaaga ccaaagcaaa ggccaaagcc aagaaaggga agggaaagga caaaggtacc
     2221 ggaggttccg gtggttccgg tctgaatgat atctttgaag ctcagaagat tgaatggcat
     2281 gaaggacgta ccaagcacca ccatcaccat cactaatgat cactcgagac tagtatcgcg
     2341 ataattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2401 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2461 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2521 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2581 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     2641 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     2701 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     2761 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     2821 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     2881 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3061 tgtcgtgcca gcggatcgat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3121 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3721 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5041 cgcgcacatt tccccgaaaa gtgccacctg ggtc
//
