LOCUS       sequence-374368-        6400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of SFRP1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 6400)
  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-Flag-hSFRP1
FEATURES             Location/Qualifiers
     source          1..6400
                     /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             664..687
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    709..733
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1676..1700)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1719..1737)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1720..1737)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1757..1774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1763..1987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2033..2461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2120..2139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2330..2351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2470..2490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2475..2804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2655..2790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2717..2736
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2871..3665
                     /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(2925..2944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3535..3554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3839..3960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3876..3895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3930..3949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4009..4025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4009..4025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4022..4044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4033..4049
                     /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(4057..4063,4064..4081,4082..4087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4102..4123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4240..4257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4411..4999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4491..4510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5170..5961,5962..6030))
                     /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     5793..5812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6031..6135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6210..6229)
                     /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 atggactaca aggacgacga tgacaagggt ggtggatccg aattatcaac aagtttgtac
      721 aaaaaagttg gcatgggcat cgggcgcagc gaggggggcc gccgcggggc agccctgggc
      781 gtgctgctgg cgctgggcgc ggcgcttctg gccgtgggct cggccagcga gtacgactac
      841 gtgagcttcc agtcggacat cggcccgtac cagagcgggc gcttctacac caagccacct
      901 cagtgcgtgg acatccccgc ggacctgcgg ctgtgccaca acgtgggcta caagaagatg
      961 gtgctgccca acctgctgga gcacgagacc atggcggagg tgaagcagca ggccagcagc
     1021 tgggtgcccc tgctcaacaa gaactgccac gccggcaccc aggtcttcct ctgctcgctc
     1081 ttcgcgcccg tctgcctgga ccggcccatc tacccgtgtc gctggctctg cgaggccgtg
     1141 cgcgactcgt gcgagccggt catgcagttc ttcggcttct actggcccga gatgcttaag
     1201 tgtgacaagt tccccgaggg ggacgtctgc atcgccatga cgccgcccaa tgccaccgaa
     1261 gcctccaagc cccaaggcac aacggtgtgt cctccctgtg acaacgagtt gaaatctgag
     1321 gccatcattg aacatctctg tgccagcgag tttgcactga ggatgaaaat aaaagaagtg
     1381 aaaaaagaaa atggcgacaa gaagattgtc cccaagaaga agaagcccct gaagttgggg
     1441 cccatcaaga agaaggacct gaagaagctt gtgctgtacc tgaagaatgg ggctgactgt
     1501 ccctgccacc agctggacaa cctcagccac cacttcctca tcatgggccg caaggtgaag
     1561 agccagtact tgctgacggc catccacaag tgggacaaga aaaacaagga gttcaaaaac
     1621 ttcatgaaga aaatgaaaaa ccatgagtgc cccacctttc agtccgtgtt taagtgccca
     1681 actttcttgt acaaagtggt tgatctagag ggccctattc tatagtgtca cctaaatgct
     1741 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1801 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1861 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1921 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1981 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2161 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2221 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2281 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2341 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2401 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2461 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2521 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2641 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2701 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     2761 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     2821 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2881 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2941 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3001 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3061 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3121 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3181 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3241 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3301 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3361 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3421 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3481 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3541 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3601 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3661 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3721 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3781 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3841 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3901 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3961 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4021 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4081 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4141 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4201 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4261 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4321 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4381 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4921 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4981 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5041 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5101 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5161 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5221 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5281 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5341 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5401 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5461 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5521 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5581 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5641 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5701 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5761 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5821 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5881 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5941 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6001 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6121 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6181 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6241 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6301 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     6361 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
