LOCUS       sequence-366640-        4797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the brine shrimp protein 
            B9VQB0_ARTSF. This is one of the bottom performing targets in our 
            apoptosis assay (it induced apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4797)
  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 4797)
  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: B9VQB0_ARTSF pSecTag2(pm) (pBS0257)
FEATURES             Location/Qualifiers
     source          1..4797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1041..1060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1232..1611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1612..1815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1766..1786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1860..1879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1895..1904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1904..1927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        2897..2944
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2963..3337
                     /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    3467..3588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3504..3523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3558..3577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3637..3653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3637..3653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3650..3672)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3661..3677
                     /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(3685..3691,3692..3709,3710..3715))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3868..3885)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4039..4627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4119..4138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     1021 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     1081 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     1141 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     1201 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     1261 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     1321 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     1381 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     1441 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     1501 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1561 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1621 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1681 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1741 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1801 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     1861 aatacgactc actataggga gacccgaatt cgccgccacc atggattaca aggatgatga
     1921 tgataagatg atgttgtcta agagattgat taagtctttg tcatgtgtgt ctagaacaga
     1981 attgagggct ttctcaggta ccacatcttg ttgtttgcaa caaaaggatt tagataagaa
     2041 caagggtgat actccaccac catctaggga acatgaggag caggagggag ttttcaagag
     2101 ggccatggag aaggccaagg gtgaatacga ccctgaatac cctttatctt ctagcatgaa
     2161 agccaccaag gatgttgcaa aggacgtggc cgaaggagct aaagagaagg ttaagtctgc
     2221 ctacgaatct attaaggaga gtgtctcttc caccagttcc gaagcacaaa acagaggtga
     2281 gtccatgtat ggtaagacca aagaaaccgt ttctgatact gctaacaagg ctaaggaaaa
     2341 ggccgaatct atgtatgata ccgcaaaaga gacagctaaa tctggtgctg ataagttgtc
     2401 ctgggaagat accaaggaaa cctacaagga aaaggctggt gaaattaagg agagaattca
     2461 ggataccgcc gagtctatga aagagagaat gggtgagacc ggtcacaata tgaaggaaaa
     2521 gatgcaacat actggacaat ctatgaaaga aggaatgaaa gaatcttggg aatctttgaa
     2581 ggatactgcc aagcaaacaa aagaaggtgc tcatgatcaa tggaatacag ctaaggataa
     2641 aaccaaagaa gtgaaagatg cagcatccga aaagatgtct aactcagttg acaagacttt
     2701 gaaaagaggt gaaaaagttt cagaaagagt tactgaaatg tactctggta ctaagggtga
     2761 ttctaaggga ggttcagggt ttaatcaaat taccccagag caaacagaaa acatgaaggg
     2821 tcaacaatct gcttctggtg ctcatgaaag ataagggtgt ttgtggtttt gttgtgtggt
     2881 attgtgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     2941 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     3001 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     3061 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     3121 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     3181 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     3241 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     3301 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     3361 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3421 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3481 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3541 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3601 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3661 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3721 gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3781 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3841 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3901 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3961 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4441 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4561 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4621 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4681 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4741 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacag
//
