LOCUS       sequence-363893-        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            MAHS_RAMVA_mts_1-81. 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 5387)
  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 5387)
  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_MAHS_RAMVA_mts_1-81 (pBS0918)
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1165..1188
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1400..1417)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1406..1630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1676..2104
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1763..1782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1973..1994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2113..2133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2118..2447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2298..2433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2360..2379
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2495..2542
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2561..2935
                     /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    3065..3186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3102..3121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3156..3175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3235..3251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3235..3251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3248..3270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3259..3275
                     /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(3283..3289,3290..3307,3308..3313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3466..3483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3637..4225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3717..3736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4396..5187,5188..5256))
                     /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     5019..5038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5257..5361)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      901 ctagccacca tgagtagata tcttctgaga gacgttcaag cagtgttaag aggggtgcgc
      961 aaagtggctg aatcctcact gaagctggag acagaaaaag tgtctcttag actgggggat
     1021 ttcagaagcc agccttccct gagatctgtg cccgcctccc tgacctcccg gtcccaggct
     1081 ttctccctgc aggagattgc cgcccgggct ggcgtggtgc tgcggggcgt gcagcagcag
     1141 ttcagaaacg tgtgaggatc tggggactac aaggacgacg acgacaagtg atgaattgcc
     1201 aattgaaatt tgacggccta tgtcactgaa actatccaaa caccccatgt cgatactgaa
     1261 cgtatcgacg catacctcct tccttgaaaa cgcacaatca tacaactggg cacataatgc
     1321 gtacgcccat ctagtacacc catctctgta ggtccagttc aagagctgga agagcaccct
     1381 ccacttaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1441 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1501 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1561 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1621 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1801 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2041 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2101 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2401 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2461 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     2521 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
     2581 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     2641 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     2701 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     2761 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     2821 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     2881 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
     2941 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3061 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3121 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3181 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
