LOCUS       sequence-366749-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the brine shrimp protein 
            A6YT97_ARTSF. 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 5495)
  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 5495)
  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_A6YT97_ARTSF (pBS0322)
FEATURES             Location/Qualifiers
     source          1..5495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        872..890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1472..1495
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1512..1529)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1518..1742
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1788..2216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1875..1894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2085..2106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2225..2245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2230..2559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2410..2545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2472..2491
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2607..2654
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2673..3047
                     /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    3177..3298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3214..3233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3268..3287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3347..3363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3347..3363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3360..3382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3371..3387
                     /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(3395..3401,3402..3419,3420..3425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3578..3595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3749..4337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3829..3848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4508..5299,5300..5368))
                     /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     5131..5150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5369..5473)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      841 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      901 ctggctagcc accatgagtg aacagggaaa gttaagcaga caggaagcgg gtcagagggg
      961 cgggcaggct agagctgaac agttagggca cgagggctat gttgaaatgg gcaggaaggg
     1021 aggtcaggcc agagctgagc aattagggca cgaagggtat caggagatgg ggcagaaggg
     1081 ggggcaagcc agagctgaac aattaggcac agagggatac caggagatgg gacagaaggg
     1141 cggacaaaag agggcagaac agttgggaca cgaggggtat gttgagatgg ggcagaaagg
     1201 cggccagacc agagctgaac agctgggaca cgaggggtat caggagatgg gacagaaagg
     1261 aggacagacc cgcgccgagc agttggggac cgaagggtac caggagatgg gccagaaagg
     1321 gggccagact agagccgagc agttggggca cgaaggctat gttaagatgg gcaaattagg
     1381 gggcgaggcc agaaagcaac agatgagtag ggctgactac gctgctatgg gacaaaaagg
     1441 cgggttagcc agacagcagt agggatctgg ggactacaag gacgacgacg acaagtgaac
     1501 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1561 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1621 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1681 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1741 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1801 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1861 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1921 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1981 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2041 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2101 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2161 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2221 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2341 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2461 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2521 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2581 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     2641 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     2701 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2761 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2821 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2881 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2941 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3001 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     3061 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3121 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3181 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3241 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3301 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3361 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3421 gtaaagcctg gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3481 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5461 taggggttcc gcgcacattt ccccgaaaag tgcca
//
