LOCUS       sequence-363964-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein SYUA_HUMAN 
            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 6889)
  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 6889)
  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: SYUA_HUMAN_mCh-2xFKBP (pBS1135)
FEATURES             Location/Qualifiers
     source          1..6889
                     /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"
     protein_bind    829..847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    850..868
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     878..902
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1011..1020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2178..2498
                     /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             2505..2825
                     /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             2832..2858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2906..2923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2912..3136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3182..3610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3269..3288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3479..3500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3619..3639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3624..3953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3804..3939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3866..3885
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4001..4048
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4067..4441
                     /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    4571..4692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4608..4627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4662..4681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4741..4757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4741..4757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4754..4776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4765..4781
                     /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(4789..4795,4796..4813,4814..4819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4834..4855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4972..4989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5143..5731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5223..5242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5902..6693,6694..6762))
                     /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     6525..6544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6763..6867)
                     /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 cctatcagtg
      841 atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga
      901 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      961 gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc gccaccatgg
     1021 acgtcttcat gaaggggctg agcaaggcga aggagggggt ggtggccgcc gccgagaaga
     1081 ccaagcaggg cgtggccgag gccgccggta aaactaaaga aggggtctta tatgtcggtt
     1141 ctaaaactaa agaaggagtt gtgcacggcg tggccaccgt ggccgagaag accaaggagc
     1201 aggtgaccaa cgtgggcggc gccgtggtga ccggcgtgac cgccgtggcc cagaagaccg
     1261 tggagggcgc cggcagcatc gccgccgcca ccggcttcgt gaagaaggac cagctgggca
     1321 agaacgagga gggcgccccc caggagggca tcctggagga tatgcccgtg gaccccgaca
     1381 acgaggccta tgagatgccc agcgaggagg gctaccagga ctacgagccc gaggcctccg
     1441 gaggcggcgg atccggcgga atggtgtcta aaggcgagga ggataacatg gctataatca
     1501 aggaatttat gaggttcaag gtgcatatgg agggatctgt gaacggtcac gagttcgaaa
     1561 tcgagggcga aggcgagggg cgaccctatg aagggacaca gacggccaaa ctgaaggtga
     1621 ccaagggtgg ccccctgcct ttcgcctggg acatcctgtc accccagttt atgtacggct
     1681 ctaaggctta cgttaagcac cctgccgata ttccggacta cttgaaactg agcttccctg
     1741 aagggttcaa gtgggaacgg gtaatgaatt ttgaggatgg cggtgtagtc acagttaccc
     1801 aggacagctc tcttcaggac ggagaattta tctataaggt taaacttcgg ggcactaact
     1861 tcccatcaga cggccccgtg atgcagaaga aaactatggg ctgggaagcc agctcagaac
     1921 gcatgtatcc cgaggacgga gcactgaagg gagaaatcaa gcagcgactc aagctgaagg
     1981 atgggggaca ttatgatgct gaggtcaaaa ccacctataa ggccaagaaa cccgttcagc
     2041 ttcctggggc atacaacgtg aatatcaaac tggatattac atcccataac gaagactata
     2101 ccatcgtgga acagtacgag cgggccgagg gcagacatag cacagggggc atggatgaat
     2161 tgtacaaggc gactagaggc gtccaagtcg aaaccattag tcccggcgat ggcagaacat
     2221 ttcctaaaag gggacaaaca tgtgtcgtcc attatacagg catgttggag gacggcaaaa
     2281 aggtggacag tagtagagat cgcaataaac ctttcaaatt catgttggga aaacaagaag
     2341 tcattagggg atgggaggag ggcgtggctc aaatgtccgt cggccaacgc gctaagctca
     2401 ccatcagccc cgactacgca tacggcgcta ccggacatcc cggaattatt ccccctcacg
     2461 ctaccttggt gtttgacgtc gaactgttga agctcgagac tagaggagtg caggtggaga
     2521 ctatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     2581 acaccgggat gcttgaagat ggaaagaaag ttgattcctc ccgggacaga aacaagccct
     2641 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     2701 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     2761 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     2821 tggaaactag ttatccgtac gacgtaccag actacgcata agcggccgct cgagtctaga
     2881 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2941 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3001 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3061 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3121 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3181 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3361 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3481 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3601 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3721 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3781 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3841 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3901 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3961 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4021 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     4081 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     4141 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     4201 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     4261 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     4321 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     4381 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     4441 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4501 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4561 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4741 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4801 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4861 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4921 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5941 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6001 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6061 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgcca
//
