LOCUS       sequence-366745-        5657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein MAHS_RAMVA. 
            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 5657)
  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 5657)
  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 (pBS0310)
FEATURES             Location/Qualifiers
     source          1..5657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..53)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        225..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        605..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     853..872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        853..871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1615..1638
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1655..1672)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1661..1885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1931..2359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2018..2037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2228..2249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2368..2388)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2373..2702
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2553..2688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2615..2634
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2750..2797
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2816..3190
                     /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    3320..3441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3357..3376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3411..3430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3490..3506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3490..3506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3503..3525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3514..3530
                     /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(3538..3544,3545..3562,3563..3568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3721..3738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3892..4480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3972..3991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4651..5442,5443..5511))
                     /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     5274..5293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5512..5616)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
       61 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
      121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      841 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc caccatgtcc
      901 agatacttac taagagacgt gcaagcagtt cttagagggg gcgtgaaggt tgctgaatca
      961 agtctaaaac tggaaacaga aaaagtcagt ctgagactgg gagattttag aagtcaacca
     1021 tcattaaggt ctgttcccgc atctctgacc agcagatctc aggcctttag tctgcaggag
     1081 atcgccgccc gcgccggcgt ggtgctgaga ggggtgcagc agcagtttag aaacgtgacc
     1141 ggcgtgaacg ccgcccccgt ggtggccttc gataacggga gcgtgctgta ctccgagagg
     1201 atccacagtc agagctctca gaagcaggcc ccaaccaccg tgccaaccgg atctgtgagc
     1261 aactcccctc agccagaggg caaggccaac gaggccgccg agagagccaa gcagttcatg
     1321 aacccccctg tggcccctat ggacccagtc gacaagaacg agttcgttgc catgcctgag
     1381 atgggccgct ccaatggcaa cggagagaac aagcaggccg ccgacttcat gaagaaccag
     1441 ggcgacaccg atatggacag ccagtacgca cccgactcca gcaagaacac caagagcgtg
     1501 cccaccaagg agatcgtggc cgaggacggg tccatgtcta tcgaggacat caagaaggcc
     1561 acccaggtga cccctggggt ggccgtgaag aacgagggcg tgtgaggatc tggggactac
     1621 aaggacgacg acgacaagtg aacccgctga tcagcctcga ctgtgccttc tagttgccag
     1681 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1741 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1801 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1861 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     1921 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1981 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2221 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2281 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2341 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2461 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2521 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2581 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2641 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2701 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     2761 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     2821 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     2881 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     2941 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     3001 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     3061 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     3121 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     3181 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3241 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3301 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3361 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3421 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3481 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3541 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaacttac
     3601 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3661 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3721 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3781 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3841 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3901 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3961 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4021 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4081 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4141 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4201 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4261 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4321 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4381 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4441 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4501 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4561 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4621 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4681 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4741 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4801 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4861 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4921 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4981 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5041 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5101 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5161 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5221 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5281 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5341 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5401 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5461 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5521 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5581 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5641 tgacgtcgac ggatcgg
//
