LOCUS       sequence-374538-        6640 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 6640)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 6640)
  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: pSCM167-Venus-VF-hSFRP1(aa181-314)
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(664..666,667..669,670..1380)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1420..1443
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1465..1489
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1916..1940)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1959..1977)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1960..1977)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1997..2014)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2003..2227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2273..2701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2360..2379)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2570..2591
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2710..2730)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2715..3044
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2895..3030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2957..2976
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3111..3905
                     /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(3165..3184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3775..3794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4079..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4170..4189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4249..4265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4249..4265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4262..4284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4273..4289
                     /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(4297..4303,4304..4321,4322..4327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4342..4363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4480..4497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4651..5239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4731..4750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5410..6201,6202..6270))
                     /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     6033..6052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6271..6375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6450..6469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1141 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 ggagggggtg gatctggagg gggtggatct aagcttatgg actacaagga cgacgatgac
     1441 aagggtggtg gatccgaatt atcaacaagt ttgtacaaaa aagcaggctt cgaaggagat
     1501 agaaccatgg cccaaggcac aacggtgtgt cctccctgtg acaacgagtt gaaatctgag
     1561 gccatcattg aacatctctg tgccagcgag tttgcactga ggatgaaaat aaaagaagtg
     1621 aaaaaagaaa atggcgacaa gaagattgtc cccaagaaga agaagcccct gaagttgggg
     1681 cccatcaaga agaaggacct gaagaagctt gtgctgtacc tgaagaatgg ggctgactgt
     1741 ccctgccacc agctggacaa cctcagccac cacttcctca tcatgggccg caaggtgaag
     1801 agccagtact tgctgacggc catccacaag tgggacaaga aaaacaagga gttcaaaaac
     1861 ttcatgaaga aaatgaaaaa ccatgagtgc cccacctttc agtccgtgtt taagaaccca
     1921 gctttcttgt acaaagtggt tgatctagag ggccctattc tatagtgtca cctaaatgct
     1981 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2041 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2101 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2161 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2221 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2281 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2341 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2401 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2461 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2521 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2581 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2641 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2701 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2761 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2821 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2881 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2941 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3001 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3061 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3121 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3181 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3241 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3301 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3361 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3421 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3481 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3541 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3601 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3661 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3721 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3781 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3841 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3901 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3961 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4021 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4201 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5521 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6421 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6481 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6541 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     6601 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
