LOCUS       sequence-419571-        6414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA tagged human p62.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  AUTHORS   Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q
  TITLE     Keap1 facilitates p62-mediated ubiquitin aggregate clearance via 
            autophagy.
  JOURNAL   Autophagy. 2010 Jul 22. 6(5):.
  PUBMED    20495340
REFERENCE   2  (bases 1 to 6414)
  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: "Autophagy. 
            2010 Jul 22. 6(5):."
COMMENT     Sequence Label: HA-p62
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     501..520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(739..843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(918..937)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1109..1488
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1489..1692
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1643..1663
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1694..1712
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1715..1733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1743..1767
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3229..3255
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3296..3313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3302..3526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3572..4000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3659..3678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3869..3890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4009..4029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4014..4343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4194..4329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4256..4275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4391..4438
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4457..4831
                     /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    4961..5082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4998..5017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5052..5071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5131..5147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5131..5147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5144..5166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5155..5171
                     /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(5179..5185,5186..5203,5204..5209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5224..5245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5362..5379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5533..6121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5613..5632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6292..6414,1..669,670..738))
                     /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"
ORIGIN
        1 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
       61 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      121 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      181 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      241 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      301 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      361 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      421 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      481 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      541 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      601 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      661 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      721 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      781 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      841 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
      901 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
      961 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     1021 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     1081 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     1141 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1201 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1261 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1321 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1381 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1441 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1501 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1561 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1621 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1681 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
     1741 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     1801 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagcttggt
     1861 accgagctcg gatccactag tccagtgtgg tggaattcta tggcgtcgct caccgtgaag
     1921 gcctaccttc tgggcaagga ggacgcggcg cgcgagattc gccgcttcag cttctgctgc
     1981 agccccgagc ctgaggcgga agccgaggct gcggcgggtc cgggaccctg cgagcggctg
     2041 ctgagccggg tggccgccct gttccccgcg ctgcggcctg gcggcttcca ggcgcactac
     2101 cgcgatgagg acggggactt ggttgccttt tccagtgacg aggaattgac aatggccatg
     2161 tcctacgtga aggatgacat cttccgaatc tacattaaag agaaaaaaga gtgccggcgg
     2221 gaccaccgcc caccgtgtgc tcaggaggcg ccccgcaaca tggtgcaccc caatgtgatc
     2281 tgcgatggct gcaatgggcc tgtggtagga acccgctaca agtgcagcgt ctgcccagac
     2341 tacgacttgt gtagcgtctg cgagggaaag ggcttgcacc gggggcacac caagctcgca
     2401 ttccccagcc ccttcgggca cctgtctgag ggcttctcgc acagccgctg gctccggaag
     2461 gtgaaacacg gacacttcgg gtggccagga tgggaaatgg gtccaccagg aaactggagc
     2521 ccacgtcctc ctcgtgcagg ggaggcccgc cctggcccca cggcagaatc agcttctggt
     2581 ccatcggagg atccgagtgt gaatttcctg aagaacgttg gggagagtgt ggcagctgcc
     2641 cttagccctc tgggcattga agttgatatc gatgtggagc acggagggaa aagaagccgc
     2701 ctgacccccg tctctccaga gagttccagc acagaggaga agagcagctc acagccaagc
     2761 agctgctgct ctgatcccag caagccgggt gggaatgttg agggcgccac gcagtctctg
     2821 gcggagcaga tgagaaagat cgccttggag tccgaggggc gccctgagga acagatggag
     2881 tcggataact gttcaggagg agatgatgac tggacccatc tgtcttcaaa agaagtggac
     2941 ccgtctacag gtgaactcca gtccctacag atgccagaat ccgaagggcc aagctctctg
     3001 gacccctccc aggagggacc cacagggctg aaggaagctg ccttgtaccc acatctcccg
     3061 ccagaggctg acccgcggct gattgagtcc ctctcccaga tgctgtccat gggcttctct
     3121 gatgaaggcg gctggctcac caggctcctg cagaccaaga actatgacat cggagcggct
     3181 ctggacacca tccagtattc aaagcatccc ccgccgttgc gcggccgcta cccatacgat
     3241 gttccagatt acgcttgact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3301 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3361 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3421 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3481 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3541 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3601 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3661 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3721 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3781 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3841 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3901 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3961 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4021 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4261 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4321 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4381 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     4441 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     4501 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     4561 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     4621 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     4681 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     4741 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     4801 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     4861 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4921 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4981 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5041 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5101 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5161 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5221 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5281 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5341 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5401 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5461 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5701 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5761 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5821 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5881 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5941 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6001 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6061 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6121 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6181 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6241 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6301 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6361 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccag
//
