LOCUS       sequence-9998-ex        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  AUTHORS   Stanhill A, Haynes CM, Zhang Y, Min G, Steele MC, Kalinina J, 
            Martinez E, Pickart CM, Kong XP, Ron D
  TITLE     An arsenite-inducible 19S regulatory particle-associated protein 
            adapts proteasomes to proteotoxicity.
  JOURNAL   Mol Cell. 2006 Sep 15. 23(6):875-85.
  PUBMED    16973439
REFERENCE   2  (bases 1 to 6454)
  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"; journalName: "Mol Cell. 
            2006 Sep 15. 23(6):875-85."
COMMENT     Sequence Label: AIRAP 5: AIRAP(1-50).GST,V1.pcDNA3
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1285..1938
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1896..1918
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1945..1962
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(2006..2024)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2007..2024)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2044..2061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2050..2274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2320..2748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2407..2426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2617..2638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2757..2777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2762..3092
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2943..3078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3005..3024
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3159..3953
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3213..3232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3823..3842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4127..4248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4164..4183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4218..4237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4297..4313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4297..4313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4310..4332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4321..4337
                     /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(4345..4351,4352..4369,4370..4375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4390..4411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4528..4545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4699..5287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4779..4798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5458..6249,6250..6318))
                     /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     6081..6100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6319..6423)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcggcacga gggccttgag
      961 agaagtcggg cgcttcgcgg cgattggagc tgcagtttct actgcggtgt caccgggata
     1021 accacaaagg gcaccgcttg tctgtctccg cctctgacta cccgtgatac ttaaatacat
     1081 ctgtgttttt ggacagcatt tataatggag tttcctgact tggggaagca ctgttcagaa
     1141 ccaacttgca aacagctaga ttttctgcca ataacatgcg acgcctgtaa acaagatttc
     1201 tgtaaagacc atttttccta cgtgggtcat aagtgtccct ttgcgttcaa gaagggatat
     1261 ccatcacact ggcggccgct cgagatgtcc cctatactag gttattggaa aattaagggc
     1321 cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg
     1381 tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt
     1441 cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata
     1501 cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt
     1561 tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt
     1621 aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg
     1681 ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac
     1741 ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg
     1801 ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac
     1861 ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt
     1921 ggcgaccatc ctccaaaatc ggatctggtt ccgcgtggat ccccgggaat ttccggtggt
     1981 ggtggtggaa tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2041 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2101 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2161 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2221 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2281 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2461 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2521 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2641 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     2701 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     2821 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2881 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2941 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3001 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3061 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3121 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3181 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3241 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3301 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3361 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3421 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3481 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3541 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3601 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3661 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3721 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3781 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3841 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3901 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3961 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4021 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4081 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4141 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4201 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4261 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4321 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4381 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4441 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4501 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4561 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4861 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     4921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5101 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5401 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5461 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5521 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5581 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5641 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5701 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5761 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5821 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5881 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5941 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6001 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6061 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6121 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6181 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6241 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6301 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6421 gcgcacattt ccccgaaaag tgccacctga cgtc
//
