LOCUS       sequence-410359-        5575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRZB protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5575)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 5575)
  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: FRZBA-c400
FEATURES             Location/Qualifiers
     source          1..5575
                     /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             2740..2784
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2797..2814
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2926..2945)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2991..3046
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3045..3064)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3407..3423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3407..3423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3420..3442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3431..3447
                     /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(3455..3461,3462..3479,3480..3485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3500..3521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3650..3667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3821..4409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3901..3920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4580..5371,5372..5440))
                     /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     5203..5222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5441..5545)
                     /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 accggtgcag cctgtgagcc cgtccgcatc
     1861 cccctgtgca agtccctgcc ctggaacatg actaagatgc ccaaccacct gcaccacagc
     1921 actcaggcca acgccatcct ggccatcgag cagttcgaag gtctgctggg cacccactgc
     1981 agccccgatc tgctcttctt cctctgtgcc atgtacgcgc ccatctgcac cattgacttc
     2041 cagcacgagc ccatcaagcc ctgtaagtct gtgtgcgagc gggcccggca gggctgtgag
     2101 cccatactca tcaagtaccg ccactcgtgg ccggagaacc tggcctgcga ggagctgcca
     2161 gtgtacgaca ggggcgtgtg catctctccc gaggccatcg ttactgcgga cggagctgat
     2221 tttcctatgg attctagtaa cggaaactgt agaggggcaa gcagtgaacg ctgtaaatgt
     2281 aagcctatta gagctacaca gaagacctat ttccggaaca attacaacta tgtcattcgg
     2341 gctaaagtta aagagataaa gactaagtgc catgatgtga ctgcagtagt ggaggtgaag
     2401 gagattctaa agtcctctct ggtaaacatt ccacgggaca ctgtcaacct ctataccagc
     2461 tctggctgcc tctgccctcc acttaatgtt aatgaggaat atatcatcat gggctatgaa
     2521 gatgaggaac gttccagatt actcttggtg gaaggctcta tagctgagaa gtggaaggat
     2581 cgactcggta aaaaagttaa gcgctgggat atgaagcttc gtcatcttgg actcagtaaa
     2641 agtgattcta gcaatagtga ttccactcag agtcagaagt ctggcaggaa ctcgaacccc
     2701 cggcaagcac gcaacggtac cggaggttcc ggtggttccg gtctgaatga tatctttgaa
     2761 gctcagaaga ttgaatggca tgaaggacgt accaagcacc accatcacca tcactaatga
     2821 tcactcgaga ctagtatcgc gataattcac tcctcaggtg caggctgcct atcagaaggt
     2881 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     2941 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3001 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3061 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3121 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     3181 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3241 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3301 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     3361 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     3421 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3481 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3541 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg
     3601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3661 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3721 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4261 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtc
//
