LOCUS       sequence-415576-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SEAP reporter vector for LAUNCHER system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  AUTHORS   Cui M, Lee S, Ban SH, Ryu JR, Shen M, Yang SH, Kim JY, Choi SK, Han 
            J, Kim Y, Han K, Lee D, Sun W, Kwon HB, Lee D
  TITLE     A single-component, light-assisted uncaging switch for 
            endoproteolytic release.
  JOURNAL   Nat Chem Biol. 2024 Mar;20(3):353-364. doi: 
            10.1038/s41589-023-01480-6. Epub 2023 Nov 16.
  PUBMED    37973890
REFERENCE   2  (bases 1 to 5693)
  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"; doi: 
            "10.1038/s41589-023-01480-6"; journalName: "Nat Chem Biol"; date: 
            "2024-03"; volume: "20"; issue: "3"; pages: "353-364"
COMMENT     Sequence Label: TRE-SEAP
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(21..39,40..62,63..81,82..104,105..123,124..146,
                     147..165,166..188,189..207,208..230,231..249,250..272,
                     273..291)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(63..81,82..104,105..123,124..146,147..165,166..188,
                     189..207,208..230,231..249,250..272,273..291,292..314,
                     315..333)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(105..123,124..146,147..165,166..188,189..207,208..230,
                     231..249,250..272,273..291,292..314,315..333,334..356,
                     357..375)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(147..165,166..188,189..207,208..230,231..249,250..272,
                     273..291,292..314,315..333,334..356,357..375,376..398,
                     399..417)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(189..207,208..230,231..249,250..272,273..291,292..314,
                     315..333,334..356,357..375,376..398,399..417,418..440,
                     441..459)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(231..249,250..272,273..291,292..314,315..333,334..356,
                     357..375,376..398,399..417,418..440,441..459,460..482,
                     483..501)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(273..291,292..314,315..333,334..356,357..375,376..398,
                     399..417,418..440,441..459,460..482,483..501,502..524,
                     525..543)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     complement(774..793)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     989..1013
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1167..2669
                     /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(2696..2715)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2697..2715)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2720..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2967..2987)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2967..2985)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3006..3022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3006..3022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3019..3041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3030..3046
                     /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(3054..3060,3061..3078,3079..3084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3099..3120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3237..3254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3407..3995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3487..3506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4166..4957,4958..5026))
                     /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     4789..4808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5027..5131)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5157..5612)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5294..5315)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5506..5525
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccctcgatcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
       61 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      121 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      181 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
      241 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
      301 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
      361 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      421 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      481 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      541 agaaaagtga aagtcgagct cggtacgcta tggcatgcat gtgtcgcgac ctgcaggccc
      601 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
      661 tgacctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      721 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      781 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      841 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      901 acgccatcag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
      961 acttcaagag ctagccctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     1021 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg
     1081 gccccggtac cgaattcaag gcctctcgag atccattgtg ctggatctgc gatctaagta
     1141 agcttcgaat cgcgaattcg cccaccatgc tgctgctgct gctgctgctg ggcctgaggc
     1201 tacagctctc cctgggcatc atcccagttg aggaggagaa cccggacttc tggaaccgcg
     1261 aggcagccga ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga
     1321 acctcatcat cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc
     1381 taaaagggca gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc
     1441 catatgtggc tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca
     1501 cagccacggc ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag
     1561 ccgcccgctt taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc
     1621 gggccaagaa agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct
     1681 cgccagccgg cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc
     1741 ctgcctcggc ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg
     1801 acattgacgt gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc
     1861 ctgagtaccc agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc
     1921 aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc
     1981 aggcttccct ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga
     2041 aatacgagat ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg
     2101 ccctgcgcct gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca
     2161 tcgaccatgg tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg
     2221 acgacgccat tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca
     2281 ctgccgacca ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct
     2341 tcgggctggc ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa
     2401 acggtccagg ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg
     2461 ggagccccga gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg
     2521 aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc
     2581 agaccttcat agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg
     2641 acctggcgcc ccccgccggc accaccgacg ccgcgcaccc gggttactct agcctcccta
     2701 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcgtccgcgg cgccaatgca
     2941 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     3001 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3061 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3121 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3241 ctcactgact cgctgcgctc ggtcgttcgg ctgcgggagc ggtatcagct cactcaaagg
     3301 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3361 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3421 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3481 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3541 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3601 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3661 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3721 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3781 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3841 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3901 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3961 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4021 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4081 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4141 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4201 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4261 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4321 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4381 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4441 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4501 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4561 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4621 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4681 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4741 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4801 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4861 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4921 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4981 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5041 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5101 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     5161 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5221 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5281 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5341 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5401 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5461 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5521 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5581 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     5641 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacga tat
//
