LOCUS       sequence-436956-        4833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SEAP reporter vector for optochemoWnt system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4833)
  AUTHORS   Lee S, Cui M, Lee D, Han K, Sun W, Lee D
  TITLE     A rationally designed optochemogenetic switch for activating 
            canonical Wnt signaling.
  JOURNAL   iScience. 2023 Feb 19;26(3):106233. doi: 10.1016/j.isci.2023.106233.
            eCollection 2023 Mar 17.
  PUBMED    36915690
REFERENCE   2  (bases 1 to 4833)
  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"; journalName: "iScience.";
            date: "2023-02-19"; volume: "26(3):106233. doi"; pages: " 
            10.1016/j.isci.2023.106233. eCollection 2023 Mar 17"
COMMENT     Sequence Label: 7xTCF-SEAP
FEATURES             Location/Qualifiers
     source          1..4833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        222..253
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             307..1809
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     primer_bind     complement(1836..1855)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1837..1855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1860..1994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1884..1903)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1938..1957
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2107..2127)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2107..2125)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2146..2162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2146..2162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2159..2181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2170..2186
                     /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(2194..2200,2201..2218,2219..2224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2239..2260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2377..2394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2547..3135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2627..2646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3306..4097,4098..4166))
                     /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     3929..3948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4167..4271)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4297..4752)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4434..4455)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4646..4665
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccctcgatcg agtttaccac aaagtgaaag tcgagctcgg tacgctatgg catgcctgca
       61 gggtaccgag ctcttacgcg agatcaaagg gggtaagatc aaagggggta agatcaaagg
      121 ggcgcgagat caaagggggt aagatcaaag ggggtaagat caaagggggt aagatcaaag
      181 gggcgcgccc gcgtgctagc ccgggctcga gatctagact ctagagggta tataatggaa
      241 gctcgaattc cagcttggca ttccggtact gttggtaaaa agcttcgaat cgcgaattcg
      301 cccaccatgc tgctgctgct gctgctgctg ggcctgaggc tacagctctc cctgggcatc
      361 atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga ggccctgggt
      421 gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat cttcctgggc
      481 gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca gaagaaggac
      541 aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc tctgtccaag
      601 acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc ctacctgtgc
      661 ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt taaccagtgc
      721 aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa agcagggaag
      781 tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg cacctacgcc
      841 cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc ccgccaggag
      901 gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt gatcctaggt
      961 ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc agatgactac
     1021 agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct ggcgaagcgc
     1081 cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct ggacccgtct
     1141 gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat ccaccgagac
     1201 tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct gctgagcagg
     1261 aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa
     1321 agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat tgagagggcg
     1381 ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca ctcccacgtc
     1441 ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc ccctggcaag
     1501 gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg ctatgtgctc
     1561 aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga gtatcggcag
     1621 cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg
     1681 cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat agcgcacgtc
     1741 atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc ccccgccggc
     1801 accaccgacg ccgcgcaccc gggttactct agcctcccta tagtgagtcg tattacgtag
     1861 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1921 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1981 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2041 tgtgggaggt tttttaattc gcgtccgcgg cgccaatgca ttgggcccgg tacccagctt
     2101 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2161 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     2221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2401 ggtcgttcgg ctgcgggagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2461 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2521 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2581 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2641 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2701 ctgtccgcct ttctcccttc gggaagcgtg gcgctttcta atagctcacg ctgtaggtat
     2761 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2821 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2881 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2941 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3001 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3061 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3361 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3421 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3601 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3661 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3721 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3781 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3841 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3901 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3961 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4021 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4081 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4141 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4261 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4321 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4381 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4441 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4501 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4561 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4621 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4681 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4741 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4801 gggcctcttc gctattacgc cagtcgacga tat
//
