LOCUS       sequence-374132-        6891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of SFRP1 in mammalian cells. Contains only Netrin
            (NTR) domain--amino acids 181-314..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6891)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 6891)
  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(aa181-314)
FEATURES             Location/Qualifiers
     source          1..6891
                     /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(1764..1788)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1820..1839)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1821..1839)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1850..1867)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1850..1866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1881)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2271..2405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2308..2327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2362..2381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2531..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3103..3460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3103..3294
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3126..3146)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3311..3446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3511..4305
                     /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(3565..3584)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4175..4194
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4369..4417
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4458..4477)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4577..4599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4723..4827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4828..4896,4897..5688)
                     /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(5046..5065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5859..6447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6348..6367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6601..6618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6692..6796)
                     /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 caaggcacaa cggtgtgtcc
     1381 tccctgtgac aacgagttga aatctgaggc catcattgaa catctctgtg ccagcgagtt
     1441 tgcactgagg atgaaaataa aagaagtgaa aaaagaaaat ggcgacaaga agattgtccc
     1501 caagaagaag aagcccctga agttggggcc catcaagaag aaggacctga agaagcttgt
     1561 gctgtacctg aagaatgggg ctgactgtcc ctgccaccag ctggacaacc tcagccacca
     1621 cttcctcatc atgggccgca aggtgaagag ccagtacttg ctgacggcca tccacaagtg
     1681 ggacaagaaa aacaaggagt tcaaaaactt catgaagaaa atgaaaaacc atgagtgccc
     1741 cacctttcag tccgtgttta agaacccagc tttcttgtac aaagtggttg atcgcgtgca
     1801 tgcgacgtca tagctctctc cctatagtga gtcgtattat aagctaggca ctggccgtcg
     1861 ttttacaacg tcgtgactgg gaaaactgct agcttgggat ctttgtgaag gaaccttact
     1921 tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata
     1981 taaaattttt aagtgtataa tgtgttaaac tagctgcata tgcttgctgc ttgagagttt
     2041 tgcttactga gtatgattta tgaaaatatt atacacagga gctagtgatt ctaattgttt
     2101 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt
     2161 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2221 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2401 ggatcgatcc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc
     2461 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2521 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     3001 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     3061 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     3121 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac
     3481 aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt
     3541 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3601 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3661 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3721 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3781 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3841 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3901 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3961 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4021 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4081 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4141 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4201 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4261 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4321 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     4381 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc
     4441 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4501 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4561 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4621 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4681 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4981 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5041 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6001 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6301 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6361 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6421 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6481 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6541 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6601 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6661 cgttggccga ttcattaatg cagagcttgc aattcgcgct ttttcaatat tattgaagca
     6721 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6781 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6841 ttatcatgac attaacctat aaaaataggc gtattacgag gccctttcac g
//
