LOCUS       sequence-366780-        5381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS1_RAMVA_108-197. 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 5381)
  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 5381)
  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_RAMVA_108-197 (pBS0652)
FEATURES             Location/Qualifiers
     source          1..5381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(41..60)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        232..611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        612..815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     766..786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     860..879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1386..1403)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1392..1616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1662..2090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1749..1768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1959..1980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2099..2119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2104..2433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2284..2419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2481..2528
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2547..2921
                     /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    3051..3172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3088..3107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3142..3161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3221..3237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3221..3237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3234..3256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3245..3261
                     /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(3269..3275,3276..3293,3294..3299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3452..3469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3623..4211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3703..3722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4382..5173,5174..5242))
                     /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     5005..5024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5243..5347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
       61 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
      121 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      181 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      481 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      541 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      601 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      721 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      781 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      841 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagccac
      901 catggagcgg tatcagaagg agcacgagat gatcaacaga caggccgaag ccacactgca
      961 gaaaaaggcc gaggagtacc ggcaccagac cgaggccgag gccgagaaaa tccgcagaga
     1021 gttggagaag cagcacgaga gggatattca gttcagaaag gacctgattg atcagactat
     1081 cgagaagcag aagagagaag tggatctgga ggctaaaatg gccaagcggg agttagacag
     1141 agaagctcag ttggcaaaag aggccttaga acgttgatga attgccaatt gaaatttgac
     1201 ggcctatgtc actgaaacta tccaaacacc ccatgtcgat actgaacgta tcgacgcata
     1261 cctccttcct tgaaaacgca caatcataca actgggcaca taatgcgtac gcccatctag
     1321 tacacccatc tctgtaggtc cagttcaaga gctggaagag caccctccac ttacccgctg
     1381 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1441 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1501 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1561 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     1621 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     1681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1801 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2101 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2161 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2221 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2281 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2341 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2401 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2461 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     2521 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     2581 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     2641 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     2701 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     2761 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     2821 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     2881 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     2941 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3001 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3061 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3121 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3181 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3241 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3301 cctggggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     3361 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3481 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3541 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3601 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3661 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3721 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3781 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3841 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3901 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3961 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4021 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4081 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4141 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5341 ttccgcgcac atttccccga aaagtgccac ctgacgtcga c
//
