LOCUS       sequence-374145-        6840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of SFRP1 in mammalian cells. Contains only 
            Frizzled cysteine rich domain (CRD)--amino acids 53-169..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 6840)
  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: pDEST27-GST-hSFRP1(aa53-169)
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             632..1285
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1243..1265
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    1313..1337
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1713..1737)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1769..1788)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1770..1788)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1799..1816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1799..1815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2220..2354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2257..2276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2311..2330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2480..2935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2567..2586)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2777..2798
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3052..3409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3052..3243
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3075..3095)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3260..3395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3460..4254
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3514..3533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4124..4143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4318..4366
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4407..4426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4526..4548
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4672..4776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4777..4845,4846..5637)
                     /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     complement(4995..5014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5808..6396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6297..6316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6550..6567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6641..6745)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggac catggcccct atactaggtt attggaaaat
      661 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
      721 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
      781 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
      841 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
      901 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
      961 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
     1021 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
     1081 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
     1141 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
     1201 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
     1261 tggtggtggc gaccatcctc caaaatcgga tctggttccg cgttctagat caacaagttt
     1321 gtacaaaaaa gcaggcttcg aaggagatag aaccatggcc accaagccac ctcagtgcgt
     1381 ggacatcccc gcggacctgc ggctgtgcca caacgtgggc tacaagaaga tggtgctgcc
     1441 caacctgctg gagcacgaga ccatggcgga ggtgaagcag caggccagca gctgggtgcc
     1501 cctgctcaac aagaactgcc acgccggcac ccaggtcttc ctctgctcgc tcttcgcgcc
     1561 cgtctgcctg gaccggccca tctacccgtg tcgctggctc tgcgaggccg tgcgcgactc
     1621 gtgcgagccg gtcatgcagt tcttcggctt ctactggccc gagatgctta agtgtgacaa
     1681 gttccccgag ggggacgtct gcatcgccat gaacccagct ttcttgtaca aagtggttga
     1741 tcgcgtgcat gcgacgtcat agctctctcc ctatagtgag tcgtattata agctaggcac
     1801 tggccgtcgt tttacaacgt cgtgactggg aaaactgcta gcttgggatc tttgtgaagg
     1861 aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta
     1921 aggtaaatat aaaattttta agtgtataat gtgttaaact agctgcatat gcttgctgct
     1981 tgagagtttt gcttactgag tatgatttat gaaaatatta tacacaggag ctagtgattc
     2041 taattgtttg tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc
     2101 acagtctgtt catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2161 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2221 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2281 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2341 atcatgtctg gatcgatcct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2401 cgtattggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2461 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2521 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2581 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2641 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2701 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2761 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2821 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2881 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     2941 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     3001 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3061 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc
     3421 ttctgacaca acagtctcga acttaaggct agagccacca tgattgaaca agatggattg
     3481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3961 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4261 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     4321 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     4381 aggatccgcg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4441 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4501 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4561 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4621 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4981 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5281 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5641 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5941 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6481 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6541 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6601 ctccccgcgc gttggccgat tcattaatgc agagcttgca attcgcgctt tttcaatatt
     6661 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6721 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6781 aaaccattat tatcatgaca ttaacctata aaaataggcg tattacgagg ccctttcacg
//
