LOCUS       sequence-363951-        7090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            DSUP_RAMVA_1-208 attached to 2xFKBP. The FKBP can be used for 
            inducible dimerization and condensate formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7090)
  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 7090)
  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: DSUP_RAMVA_1-208_mCh-2xFKBP (pBS1116)
FEATURES             Location/Qualifiers
     source          1..7090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..80)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        252..631
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        632..835
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     786..806
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    858..876
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     886..910
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2387..2707
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2714..3034
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3041..3067
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3115..3132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3121..3345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3391..3819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3478..3497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3688..3709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3828..3848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3833..4162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4075..4094
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4210..4257
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4276..4650
                     /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    4780..4901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4817..4836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4871..4890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4950..4966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4950..4966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4963..4985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4974..4990
                     /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(4998..5004,5005..5022,5023..5028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5043..5064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5181..5198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5352..5940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5432..5451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6111..6902,6903..6971))
                     /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     6734..6753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6972..7076)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
       61 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
      121 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
      181 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      241 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      301 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      361 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      421 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      481 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      541 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      601 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      661 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      721 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      781 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc
      841 tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc gtttagtgaa
      901 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      961 ccgatccagc ctccggactc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc
     1021 caccatggcc tctactcatc aatcttctac tgaacctagc tccaccggca aatctgaaga
     1081 gactaagaag gacgcttccc agggcagtgg ccaggacagc aaaaacgtta ccgttaccaa
     1141 gggcaccggc tcctccgcca ccagcgccgc catcgtgaaa accggcggga gccagggcaa
     1201 ggactcaagt acaacggcgg gcagctccag cacacagggc cagaagttct ccaccacacc
     1261 aaccgaccca aagaccttca gtagtgacca gaaggagaag agcaaatcac ccgccaaaga
     1321 ggtgccctcc ggcggcgact ctaaatccca gggcgacacc aagtctcagt ctgatgccaa
     1381 gagcagcgga cagagccagg gccagagtaa ggattccggg aagtcatcct ccgacagctc
     1441 caagtctcac agcgtgatcg gcgccgtgaa agacgtggtc gccggcgcca aggacgtggc
     1501 cggcaaagcc gtggaggacg ccccttccat tatgcacacc gccgtcgatg cagtcaagaa
     1561 cgccgccaca acagtcaagg acgtggcctc tagcgcagcc agtaccgtgg ccgagaaggt
     1621 ggtggatgcc tatcattcag tggtttccgg aggcggcgga tccggcggaa tggtgtctaa
     1681 aggcgaggag gataacatgg ctataatcaa ggaatttatg aggttcaagg tgcatatgga
     1741 gggatctgtg aacggtcacg agttcgaaat cgagggcgaa ggcgaggggc gaccctatga
     1801 agggacacag acggccaaac tgaaggtgac caagggtggc cccctgcctt tcgcctggga
     1861 catcctgtca ccccagttta tgtacggctc taaggcttac gttaagcacc ctgccgatat
     1921 tccggactac ttgaaactga gcttccctga agggttcaag tgggaacggg taatgaattt
     1981 tgaggatggc ggtgtagtca cagttaccca ggacagctct cttcaggacg gagaatttat
     2041 ctataaggtt aaacttcggg gcactaactt cccatcagac ggccccgtga tgcagaagaa
     2101 aactatgggc tgggaagcca gctcagaacg catgtatccc gaggacggag cactgaaggg
     2161 agaaatcaag cagcgactca agctgaagga tgggggacat tatgatgctg aggtcaaaac
     2221 cacctataag gccaagaaac ccgttcagct tcctggggca tacaacgtga atatcaaact
     2281 ggatattaca tcccataacg aagactatac catcgtggaa cagtacgagc gggccgaggg
     2341 cagacatagc acagggggca tggatgaatt gtacaaggcg actagaggcg tccaagtcga
     2401 aaccattagt cccggcgatg gcagaacatt tcctaaaagg ggacaaacat gtgtcgtcca
     2461 ttatacaggc atgttggagg acggcaaaaa ggtggacagt agtagagatc gcaataaacc
     2521 tttcaaattc atgttgggaa aacaagaagt cattagggga tgggaggagg gcgtggctca
     2581 aatgtccgtc ggccaacgcg ctaagctcac catcagcccc gactacgcat acggcgctac
     2641 cggacatccc ggaattattc cccctcacgc taccttggtg tttgacgtcg aactgttgaa
     2701 gctcgagact agaggagtgc aggtggagac tatctcccca ggagacgggc gcaccttccc
     2761 caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaagt
     2821 tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat
     2881 ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat
     2941 atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac
     3001 tctcgtcttc gatgtggagc ttctaaaact ggaaactagt tatccgtacg acgtaccaga
     3061 ctacgcataa gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     3121 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3181 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3241 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3301 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3361 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3781 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4201 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4261 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     4321 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     4381 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     4441 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     4501 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     4561 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     4621 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     4681 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4741 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4801 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4861 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4921 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4981 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5041 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5101 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5161 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5221 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5281 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5341 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5401 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5461 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5521 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5581 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5641 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5701 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5761 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5821 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5881 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5941 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6001 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6061 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6121 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6181 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6241 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6301 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6361 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6421 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6481 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6541 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6601 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6661 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6721 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6781 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6841 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6901 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6961 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7021 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7081 tttccccgaa
//
