LOCUS       sequence-366738-        5780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the synthetic protein Baker_TJ_190. 
            This is one of the top performing targets in our apoptosis assay (it
            protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5780)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 5780)
  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: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: pST_Baker_TJ_190 (pBS0298)
FEATURES             Location/Qualifiers
     source          1..5780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..42)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        214..593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        594..797
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     748..768
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     842..861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        842..860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      878..887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1727..1750
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1767..1784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1773..1997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2043..2471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2130..2149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2340..2361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2480..2500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2485..2814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2665..2800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2727..2746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2862..2909
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2928..3302
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3432..3553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3469..3488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3523..3542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3602..3618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3602..3618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3615..3637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3626..3642
                     /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(3650..3656,3657..3674,3675..3680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3833..3850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4004..4592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4084..4103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4763..5554,5555..5623))
                     /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     5386..5405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5624..5728)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
       61 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
      121 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
      181 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
      241 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      301 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      361 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      421 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      481 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      541 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      601 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      661 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      721 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      781 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
      841 ttaatacgac tcactatagg gagacccaag ctggctagcc accatgggtg gcagcggatg
      901 gggacctctg agggaagtgt tggaagaggc ccttcgcgcg cttgaagaac ttgctcggga
      961 gcgaggtgac aaagagaaag ttgaagaggt caagaaagaa aaggaagagg aagaggaaga
     1021 aaaggaaaag aataaagaag aggcgaagga gttggcagaa cttatcaagg aaaatcctct
     1081 ggtacgcgta ttggttgagg ccctcagagc ccttgttgaa cttgctgagg agcgtgggga
     1141 tctcgagaaa gtggaagagg tggccgaaga agtgaaagaa gagatagaag agcttaggga
     1201 aaataaagaa gaagcgaaag aactcgctga acttatcaag gagaaccccc tcgttcgggt
     1261 cctcgttgag gcactgaggg cgctcgttga acttgccgaa gaacgcggtg acttggaaaa
     1321 ggtagaggaa gttgccgaag aagttaaaga agaaatcgaa gaattgagag agaacaaaga
     1381 agaagctaaa gagctcgccg agctcattaa ggaaaaccca ctcgtcaggg ttcttgtaga
     1441 ggcgctccgc gcacttgttg aattggctga agagcgcgga gaccttgaaa aggtcgaaga
     1501 agtcgcagaa gaagttaagg aagaaattga agagctcagg gagaataagg aagaggccaa
     1561 agaattggct gaattgatca aagaaaatcc gcttgtacgg gttcttgtag aagccttgcg
     1621 tgcacttgtc gaactcgccg aagaacgggg cgatctcgaa aaggttgagg aagttgcaga
     1681 agaagttaaa gaagagatag aggagcttcg ggaataagga tctggggact acaaggacga
     1741 cgacgacaag tgaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1801 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1861 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1921 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1981 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2041 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2101 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2161 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2221 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2281 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2341 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2401 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2461 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2641 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2701 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2761 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2821 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     2881 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     2941 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     3001 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     3061 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     3121 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     3181 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     3241 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     3301 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3361 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3421 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3481 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3541 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactt acattaattg
     3721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5761 acggatcggg agatctcccg
//
