LOCUS       sequence-223829-        6861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the extracellular domain of SFRP2 with N-terminal FLAG
            tag and C-terminal hIgG tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6861)
  TITLE     Comoletti Lab Plasmids
REFERENCE   2  (bases 1 to 6861)
  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: pCMV6-XL4 FLAG-SFRP2-Fc
FEATURES             Location/Qualifiers
     source          1..6861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             956..1045
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1046..1069
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1895..1918
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3227..3849
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3873..3893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3878..4207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4245..4264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4246..4264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4278..4294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4278..4294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4291..4313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4302..4318
                     /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(4326..4332,4333..4350,4351..4356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4371..4392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4509..4526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4680..5268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4760..4779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5439..6230,6231..6299))
                     /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     6062..6081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6300..6404)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6431..6859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6518..6537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6728..6749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatatcg ccaccatgga
      961 cagcaaaggt tcgtcgcaga aagggtcccg cctgctcctg ctgctggtgg tgtcaaatct
     1021 actcttgtgc cagggtgtgg tctccgacta caaagacgat gacgacaagg cggccgcttt
     1081 attcctgttc ggccagcccg acttcagcta caagagaagc aactgcaagc ccatccccgc
     1141 caacctgcag ctgtgccacg gcatcgagta ccagaacatg agactgccca acctgctggg
     1201 ccacgaaacc atgaaggagg tgctggagca ggccggcgcc tggatccccc tggtgatgaa
     1261 gcagtgccac cccgacacca agaagttcct gtgcagcctg ttcgcccctg tgtgcctgga
     1321 cgacctggac gagacaatcc agccttgcca cagcctgtgc gtccaggtga aggacagatg
     1381 cgcccccgtg atgagcgcct tcggcttccc ctggcccgac atgctggagt gcgacagatt
     1441 cccccaggac aacgatctgt gtatccccct ggccagcagc gaccatctgc tgcctgccac
     1501 cgaggaggcc cccaaggtgt gcgaggcctg caagaacaag aacgacgacg acaacgacat
     1561 catggagaca ctgtgcaaga acgacttcgc cctgaagatc aaggtgaagg agatcaccta
     1621 catcaacaga gacaccaaga tcatcctgga gacaaagagc aagaccatct acaagctgaa
     1681 cggcgtgagc gagagggacc tgaagaagag cgtgctgtgg ctgaaggaca gcctgcagtg
     1741 cacatgcgag gagatgaacg acatcaacgc cccctacctg gtgatgggcc agaagcaggg
     1801 cggcgagctg gtgatcacca gcgtgaagag atggcagaag ggccagagag agttcaagag
     1861 aatcagcaga agcatcagaa agctgcagtg cggtctagag gtgctcttcc aggggcctga
     1921 tcccgatccc gagggtgagt actaagcttc agcgctcctg cctggacgca tcccggctat
     1981 gcagccccag tccagggcag caaggcaggc cccgtctgcc tcttcacccg gaggcctctg
     2041 cccgccccac tcatgctcag ggagagggtc ttctggcttt ttccccaggc tctgggcagg
     2101 cacaggctag gtgcccctaa cccaggccct gcacacaaag gggcaggtgc tgggctcaga
     2161 cctgccaaga gccatatccg ggaggaccct gcccctgacc taagcccacc ccaaaggcca
     2221 aactctccac tccctcagct cggacacctt ctctcctccc agattccagt aactcccaat
     2281 cttctctctg cagagcccaa atcttgtgac aaaactcaca catgcccacc gtgcccaggt
     2341 aagccagccc aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc
     2401 atccagggac aggccccagc cgggtgctga cacgtccacc tccatctctt cctcagcacc
     2461 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     2521 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     2581 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     2641 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     2701 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     2761 cgagaaaacc atctccaaag ccaaaggtgg gacccgtggg gtgcgagggc cacatggaca
     2821 gaggccggct cggcccaccc tctgccctga gagtgaccgc tgtaccaacc tctgtcccta
     2881 cagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggat gagctgacca
     2941 agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg
     3001 agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact
     3061 ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg
     3121 ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac acgcagaaga
     3181 gcctctccct gtctccgggt aaatgagtgc gacggccgcg actctagggt ggcatccctg
     3241 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3301 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3361 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3421 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3481 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3541 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3601 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3661 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3721 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3781 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3841 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     4261 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4321 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4381 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4441 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5041 aacaggatta gcagagcgag gtatgtaggc ggtgctacat agttcttgaa gtggtggcct
     5101 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6421 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6481 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6541 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6601 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6661 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6721 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6781 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6841 caaaaattta acgcgaattt t
//
