LOCUS       sequence-215523-        7225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Creation of a stable cell line expressing APP mutation (AD in vitro 
            study).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7225)
  TITLE     Radenovic Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7225)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pc DNA FRT TO- APP695
FEATURES             Location/Qualifiers
     source          1..7225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             complement(1355..1366)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             1846..1866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(3177..3194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3183..3407
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3540..3559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3691..3738
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3746..4768
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3746..4768
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4898..5019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4935..4954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4989..5008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5068..5084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5068..5084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5081..5103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5092..5108
                     /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(5116..5122,5123..5140,5141..5146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5161..5182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5299..5316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5470..6058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5550..5569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6229..7020,7021..7089))
                     /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     6852..6871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7090..7194)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcatg ctgcccggtt tggcactgct cctgctggcc gcctggacgg ctcgggcgct
     1081 ggaggtaccc actgatggta atgctggcct gctggctgaa ccccagattg ccatgttctg
     1141 tggcagactg aacatgcaca tgaatgtcca gaatgggaag tgggattcag atccatcagg
     1201 gaccaaaacc tgcattgata ccaaggaagg catcctgcag tattgccaag aagtctaccc
     1261 tgaactgcag atcaccaatg tggtagaagc caaccaacca gtgaccatcc agaactggtg
     1321 caagcggggc cgcaagcagt gcaagaccca tccccacttt gtgattccct accgctgctt
     1381 agttggtgag tttgtaagtg atgcccttct cgttcctgac aagtgcaaat tcttacacca
     1441 ggagaggatg gatgtttgcg aaactcatct tcactggcac accgtcgcca aagagacatg
     1501 cagtgagaag agtaccaact tgcatgacta cggcatgttg ctgccctgcg gaattgacaa
     1561 gttccgaggg gtagagtttg tgtgttgccc actggctgaa gaaagtgaca atgtggattc
     1621 tgctgatgcg gaggaggatg actcggatgt ctggtggggc ggagcagaca cagactatgc
     1681 agatgggagt gaagacaaag tagtagaagt agcagaggag gaagaagtgg ctgaggtgga
     1741 agaagaagaa gccgatgatg acgaggacga tgaggatggt gatgaggtag aggaagaggc
     1801 tgaggaaccc tacgaagaag ccacagagag aaccaccagc attgccacca ccaccaccac
     1861 caccacagag tctgtggaag aggtggttcg agttcctaca acagcagcca gtacccctga
     1921 tgccgttgac aagtatctcg agacacctgg ggatgagaat gaacatgccc atttccagaa
     1981 agccaaagag aggcttgagg ccaagcaccg agagagaatg tcccaggtca tgagagaatg
     2041 ggaagaggca gaacgtcaag caaagaactt gcctaaagct gataagaagg cagttatcca
     2101 gcatttccag gagaaagtgg aatctttgga acaggaagca gccaacgaga gacagcagct
     2161 ggtggagaca cacatggcca gagtggaagc catgctcaat gaccgccgcc gcctggccct
     2221 ggagaactac atcaccgctc tgcaggctgt tcctcctcgg cctcgtcacg tgttcaatat
     2281 gctaaagaag tatgtccgcg cagaacagaa ggacagacag cacaccctaa agcatttcga
     2341 gcatgtgcgc atggtggatc ccaagaaagc cgctcagatc cggtcccagg ttatgacaca
     2401 cctccgtgtg atttatgagc gcatgaatca gtctctctcc ctgctctaca acgtgcctgc
     2461 agtggccgag gagattcagg atgaagttga tgagctgctt cagaaagagc aaaactattc
     2521 agatgacgtc ttggccaaca tgattagtga accaaggatc agttacggaa acgatgctct
     2581 catgccatct ttgaccgaaa cgaaaaccac cgtggagctc cttcccgtga atggagagtt
     2641 cagcctggac gatctccagc cgtggcattc ttttggggct gactctgtgc cagccaacac
     2701 agaaaacgaa gttgagcctg ttgatgcccg ccctgctgcc gaccgaggac tgaccactcg
     2761 accaggttct gggttgacaa atatcaagac ggaggagatc tctgaagtga agatggatgc
     2821 agaattccga catgactcag gatatgaagt tcatcatcaa aaattggtgt tctttgcaga
     2881 agatgtgggt tcaaacaaag gtgcaatcat tggactcatg gtgggcggtg ttgtcatagc
     2941 gacagtgatc gtcatcacct tggtgatgct gaagaagaaa cagtacacat ccattcatca
     3001 tggtgtggtg gaggttgacg ccgctgtcac cccagaggag cgccacctgt ccaagatgca
     3061 gcagaacggc tacgaaaatc caacctacaa gttctttgag cagatgcaga actagagata
     3121 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3181 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3241 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3301 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3361 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3421 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3661 acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct
     3721 agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3781 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     3841 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     3901 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     3961 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     4021 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4081 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     4141 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4201 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4261 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4321 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4381 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4441 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4501 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     4561 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4621 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4681 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4741 cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc
     4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5041 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5101 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7201 tccccgaaaa gtgccacctg acgtc
//
