LOCUS       sequence-371928-        5231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS1_PARRC_150-189. 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 5231)
  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 5231)
  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_CAHS1_PARRC_150-189 (pBS0658)
FEATURES             Location/Qualifiers
     source          1..5231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(113..133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        118..447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      298..433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     360..379
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        495..542
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             561..935
                     /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    1065..1186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1102..1121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1156..1175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1235..1251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1235..1251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1248..1270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1259..1275
                     /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(1283..1289,1290..1307,1308..1313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1466..1483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1637..2225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1717..1736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2396..3187,3188..3256))
                     /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     3019..3038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3257..3361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3436..3455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3627..4006
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4007..4210
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4161..4181
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4255..4274
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4255..4273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4631..4648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4637..4861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(4907..5231,1..104)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4994..5013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5204..5225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
       61 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
      121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      361 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
      421 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
      481 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
      541 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
      601 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
      661 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
      721 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
      781 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
      841 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
      901 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
      961 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     1021 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     1081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     1201 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1261 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1321 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1441 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2461 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2581 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2821 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2881 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2941 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3001 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3061 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3121 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3181 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3241 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3301 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3361 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     3421 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     3481 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     3541 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     3601 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     3661 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     3721 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     3781 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     3841 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     3901 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     3961 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     4021 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     4081 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     4141 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     4201 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     4261 gactcactat agggagaccc aagctggcta gccaccatgc gcaaagatat tgtagagatg
     4321 gcaatcgaaa accagaaaaa aatgattgat gtcgaaagcc gttatgcaaa aaaagatatg
     4381 gaccgcgaac gtgtgaaagt ccgtatgatg ctcgagcagt gatgaattgc caattgaaat
     4441 ttgacggcct atgtcactga aactatccaa acaccccatg tcgatactga acgtatcgac
     4501 gcatacctcc ttccttgaaa acgcacaatc atacaactgg gcacataatg cgtacgccca
     4561 tctagtacac ccatctctgt aggtccagtt caagagctgg aagagcaccc tccacttacc
     4621 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4681 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4741 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4801 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4861 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4981 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5041 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5101 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5221 actggaacaa c
//
