LOCUS       sequence-334947-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secreted alkaline phosphatase (SEAP) expression in mammalian cells. 
            Reporter plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Kaberniuk AA, Baloban M, Monakhov MV, Shcherbakova DM, Verkhusha VV
  TITLE     Single-component near-infrared optogenetic systems for gene 
            transcription regulation.
  JOURNAL   Nat Commun. 2021 Jun 23;12(1):3859. doi: 10.1038/s41467-021-24212-7.
  PUBMED    34162879
REFERENCE   2  (bases 1 to 5631)
  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: "Nat 
            Commun."; date: "2021-06-23"; volume: "12(1):3859. doi"; pages: " 
            10.1038/s41467-021-24212-7"
COMMENT     Sequence Label: pG12-SEAP
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             314..1816
                     /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"
     intron          2463..2528
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2658..2678
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3103..3237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3140..3159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3194..3213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3247..3266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3248..3266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3353..3370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3524..4112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3604..3623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4283..5074,5075..5143))
                     /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     4906..4925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5144..5248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5316..5334
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5372..5394)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5494..5513
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5594..5612
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5594..5611
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagtcg
       61 actctagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
      121 cgagtcgact ctagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac
      181 tgtcctccga gtcgactcta gcggagtact gtcctccgag cggagtactg tcctccgagc
      241 ggagtactgt cctccggcta gtagcggaga ctctagaggg tatataatgg atccccgggt
      301 accggtcgcc accatgctgc tgctgctgct gctgctgggc ctgaggctac agctctccct
      361 gggcatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
      421 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
      481 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
      541 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
      601 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
      661 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
      721 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
      781 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
      841 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
      901 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
      961 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     1021 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     1081 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     1141 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     1201 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     1261 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     1321 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     1381 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     1441 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     1501 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     1561 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     1621 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     1681 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     1741 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     1801 cgccggcacc accgacgccg cgcacccggg ttaagcggcc gcgactctag aattccgtat
     1861 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     1921 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     1981 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2041 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     2101 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     2161 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     2221 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     2281 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta
     2341 cgcctgaata agtgataata agcggatgaa tggcagaaat tcgccggatc tttgtgaagg
     2401 aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta
     2461 aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     2521 tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg
     2581 aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact
     2641 ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt
     2701 cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg
     2761 ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt
     2821 ctgtaacctt tataagtagg cataacagtt ataatcataa catactgttt tttcttactc
     2881 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
     2941 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
     3001 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3061 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3121 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3181 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctgc
     3241 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     3301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5281 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5341 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5401 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5461 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5521 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     5581 tcatacacat acgatttagg tgacactata gaactcgagc agctgaagct t
//
