LOCUS       sequence-10001-e        7244 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 7244)
  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 7244)
  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 8: pCEFL.PSMB2.GST
FEATURES             Location/Qualifiers
     source          1..7244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..1205
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          254..1196
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1153..1173
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1867..2520
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2478..2500
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2527..2544
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(2588..2606)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2589..2606)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2626..2643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2632..2856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2902..3330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2989..3008)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3199..3220
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3339..3359)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3344..3674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3525..3660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3587..3606
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3741..4535
                     /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(3795..3814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4405..4424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4709..4830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4746..4765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4800..4819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4879..4895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4879..4895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4892..4914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4903..4919
                     /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(4927..4933,4934..4951,4952..4957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5110..5127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7080..7099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
       61 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      121 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      181 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      241 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      301 tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc
      361 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      421 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      481 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      541 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      601 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      661 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      721 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      781 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      841 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      901 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
      961 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1021 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1081 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1141 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1201 cgtgaggaat tggcgaagct tcccggcacc atggagtacc tcattggcat ccagggccca
     1261 gactatgtcc tcgtcgcttc cgaccgagtg gccgccagca atattgtcca gatgaaggac
     1321 gatcatgaca agatgtttaa gatgagtgaa aaaatcttac tcctatgtgt tggagaggct
     1381 ggagacactg tacagtttgc agaatatatc cagaaaaacg tgcaacttta taaaatgcgg
     1441 aatggatatg aattgtctcc cacagcagca gctaacttca cacggcggaa cctggctgac
     1501 tgtcttcgga gtcggacccc atatcatgtg aacctcctcc tggctggcta tgacgagcat
     1561 gaagggccag ccctttacta catggactac ctggcagcct tggctaaggc tccttttgca
     1621 gctcatggct atggtgcctt tctgactctc agcatccttg accgatacta cacacctacc
     1681 atctcacgtg agagggcagt ggagcttctt aggaagtgtc tggaggagct ccagaagcgc
     1741 ttcatcttga atctgcccac cttcagtgtc cgggtcattg acaaagatgg catccacaat
     1801 ctggagaaca ttgccttccc taagcgggac tccggacgat atccatcaca ctggcggccg
     1861 ctcgagatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     1921 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     1981 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     2041 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     2101 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     2161 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     2221 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     2281 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     2341 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     2401 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     2461 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     2521 tcggatctgg ttccgcgtgg atccccggga atttccggtg gtggtggtgg aatctagagg
     2581 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2641 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2701 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2761 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2821 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2881 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2941 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3001 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     3061 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3121 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3181 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3241 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     3301 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3361 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3481 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3601 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3661 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3901 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4261 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7081 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7141 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7201 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcg
//
