LOCUS       sequence-267655-        6433 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 6433)
  AUTHORS   Yao T, Song L, Xu W, DeMartino GN, Florens L, Swanson SK, Washburn 
            MP, Conaway RC, Conaway JW, Cohen RE
  TITLE     Proteasome recruitment and activation of the Uch37 deubiquitinating 
            enzyme by Adrm1.
  JOURNAL   Nat Cell Biol. 2006 Sep . 8(9):994-1002.
  PUBMED    16906146
REFERENCE   2  (bases 1 to 6433)
  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: "Nat Cell 
            Biol. 2006 Sep . 8(9):994-1002."
COMMENT     Sequence Label: pcDNA3-HA-Uch37
FEATURES             Location/Qualifiers
     source          1..6433
                     /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"
     regulatory      908..917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             920..946
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             950..976
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1986..2004)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1987..2004)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2024..2041)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2030..2254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2300..2728
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2387..2406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2597..2618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2737..2757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2742..3071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2922..3057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2984..3003
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3138..3932
                     /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(3192..3211)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3802..3821
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4106..4227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4143..4162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4197..4216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4276..4292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4276..4292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4289..4311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4300..4316
                     /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(4324..4330,4331..4348,4349..4354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4369..4390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4507..4524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4678..5266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4758..4777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5437..6228,6229..6297))
                     /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     6060..6079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6298..6402)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctagccgcc accatggctt acccatacga tgttcctgac tatgcgggct atccctatga
      961 cgtcccggac tatgcaggat ccatgacggg caacgccggg gagtggtgcc tcatggaaag
     1021 cgaccccggg gtcttcaccg agctcattaa aggattcggt tgccgaggag cccaagtaga
     1081 agaaatatgg agtttagagc ctgagaattt tgaaaaatta aagccagttc atgggttaat
     1141 ttttcttttc aagtggcagc caggagaaga accagcaggc tctgtggttc aggactcccg
     1201 acttgacacg atattttttg ctaagcaggt aattaataat gcttgtgcta ctcaagccat
     1261 agtgagtgtg ttactgaact gtacccacca ggatgtccat ttaggcgaga cattatcaga
     1321 gtttaaagaa ttttcacaaa gttttgatgc agctatgaaa ggcttggcac tgagcaattc
     1381 agatgtgatt cgacaagtac acaacagttt cgccagacag caaatgtttg aatttgatac
     1441 gaagacatca gcaaaagaag aagatgcttt tcactttgtc agttatgttc ctgttaatgg
     1501 gagactgtat gaattagatg gattaagaga aggaccgatt gatttaggtg catgcaatca
     1561 agatgattgg atcagtgcag taaggcctgt catagaaaaa aggatacaaa agtacagtga
     1621 aggtgaaatt cgatttaatt taatggccat tgtgtctgac agaaaaatga tatatgagca
     1681 gaagatagca gagttacaaa gacaacttgc agaggaggaa cccatggata cagatcaagg
     1741 taatagtatg ttaagtgcta ttcagtcaga agttgccaaa aatcagatgc ttattgaaga
     1801 ggaagtacag aaattaaaaa gatacaagat tgagaatatc agaaggaagc ataattatct
     1861 gcctttcatt atggaattgt taaagacttt agcagaacac cagcagttaa taccactagt
     1921 agaaaaggca aaagaaaaac agaacgcaaa gaaagctcag gaaaccaaat gaattcgggc
     1981 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2041 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2101 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2161 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2221 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2281 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2341 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2401 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2461 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2521 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2581 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2641 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2701 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2821 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2941 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3001 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3061 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3121 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3181 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3241 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3301 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3361 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3421 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3481 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3541 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3601 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3661 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3721 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3781 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3841 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3901 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3961 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4261 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5101 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6421 gccacctgac gtc
//
