LOCUS       sequence-374149-        6765 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 6765)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 6765)
  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: pDEST26-his-hSFRP1
FEATURES             Location/Qualifiers
     source          1..6765
                     /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             644..661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    671..695
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1638..1662)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1694..1713)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1695..1713)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1724..1741)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1724..1740)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1733..1755)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2145..2279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2182..2201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2236..2255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2405..2860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2492..2511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2702..2723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2977..3334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2977..3168
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3000..3020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3185..3320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3385..4179
                     /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(3439..3458)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4049..4068
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4243..4291
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4451..4473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4597..4701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4702..4770,4771..5562)
                     /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(4920..4939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5733..6321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6222..6241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6475..6492
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6566..6670)
                     /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 catggcgtac taccatcacc atcaccatca
      661 ctctagatca acaagtttgt acaaaaaagt tggcatgggc atcgggcgca gcgagggggg
      721 ccgccgcggg gcagccctgg gcgtgctgct ggcgctgggc gcggcgcttc tggccgtggg
      781 ctcggccagc gagtacgact acgtgagctt ccagtcggac atcggcccgt accagagcgg
      841 gcgcttctac accaagccac ctcagtgcgt ggacatcccc gcggacctgc ggctgtgcca
      901 caacgtgggc tacaagaaga tggtgctgcc caacctgctg gagcacgaga ccatggcgga
      961 ggtgaagcag caggccagca gctgggtgcc cctgctcaac aagaactgcc acgccggcac
     1021 ccaggtcttc ctctgctcgc tcttcgcgcc cgtctgcctg gaccggccca tctacccgtg
     1081 tcgctggctc tgcgaggccg tgcgcgactc gtgcgagccg gtcatgcagt tcttcggctt
     1141 ctactggccc gagatgctta agtgtgacaa gttccccgag ggggacgtct gcatcgccat
     1201 gacgccgccc aatgccaccg aagcctccaa gccccaaggc acaacggtgt gtcctccctg
     1261 tgacaacgag ttgaaatctg aggccatcat tgaacatctc tgtgccagcg agtttgcact
     1321 gaggatgaaa ataaaagaag tgaaaaaaga aaatggcgac aagaagattg tccccaagaa
     1381 gaagaagccc ctgaagttgg ggcccatcaa gaagaaggac ctgaagaagc ttgtgctgta
     1441 cctgaagaat ggggctgact gtccctgcca ccagctggac aacctcagcc accacttcct
     1501 catcatgggc cgcaaggtga agagccagta cttgctgacg gccatccaca agtgggacaa
     1561 gaaaaacaag gagttcaaaa acttcatgaa gaaaatgaaa aaccatgagt gccccacctt
     1621 tcagtccgtg tttaagtgcc caactttctt gtacaaagtg gttgatcgcg tgcatgcgac
     1681 gtcatagctc tctccctata gtgagtcgta ttataagcta ggcactggcc gtcgttttac
     1741 aacgtcgtga ctgggaaaac tgctagcttg ggatctttgt gaaggaacct tacttctgtg
     1801 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
     1861 ttttaagtgt ataatgtgtt aaactagctg catatgcttg ctgcttgaga gttttgctta
     1921 ctgagtatga tttatgaaaa tattatacac aggagctagt gattctaatt gtttgtgtat
     1981 tttagattca cagtcccaag gctcatttca ggcccctcag tcctcacagt ctgttcatga
     2041 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2101 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     2281 atcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggctggcgt
     2341 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2401 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2461 accgctacaa ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2821 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     2881 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     2941 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3241 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3301 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     3361 ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg
     3421 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3481 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3541 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3601 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3661 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3721 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3781 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3841 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3901 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3961 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4021 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4081 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4141 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4201 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     4261 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg
     4321 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4501 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4561 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5401 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6421 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6481 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6541 ccgattcatt aatgcagagc ttgcaattcg cgctttttca atattattga agcatttatc
     6601 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6661 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6721 tgacattaac ctataaaaat aggcgtatta cgaggccctt tcacg
//
