LOCUS       sequence-273785-        6751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagged human AO7 (aa 2-439) for expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6751)
  AUTHORS   Li S, Liang YH, Mariano J, Metzger MB, Stringer DK, Hristova VA, Li 
            J, Randazzo PA, Tsai YC, Ji X, Weissman AM
  TITLE     Insights into Ubiquitination from the Unique Clamp-like Binding of 
            the RING E3 AO7 to the E2 UbcH5B.
  JOURNAL   J Biol Chem. 2015 Dec 18;290(51):30225-39. doi: 
            10.1074/jbc.M115.685867. Epub 2015 Oct 16.
  PUBMED    26475854
REFERENCE   2  (bases 1 to 6751)
  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: 
            "10.1074/jbc.M115.685867"; journalName: "J Biol Chem"; date: 
            "2015-12-18- 18"; volume: "290"; issue: "51"; pages: "30225-39"
COMMENT     Sequence Label: pCDNA3.1+  HA-hAO7
FEATURES             Location/Qualifiers
     source          1..6751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             727..753
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2108..2125)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2114..2338
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2384..2812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2471..2490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2681..2702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2821..2841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2826..3155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3006..3141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3068..3087
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3222..4016
                     /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(3276..3295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3886..3905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4190..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4227..4246)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4281..4300
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4360..4376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4360..4376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4373..4395)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4384..4400
                     /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(4408..4414,4415..4432,4433..4438))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4453..4474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4591..4608)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4762..5350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4842..4861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5521..6312,6313..6381))
                     /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     6144..6163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6382..6486)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6561..6580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcatgtacc catacgatgt tccagattac gctgcggcgt ctgcgtctgc agctgcaggg
      781 gaggaggact gggtccttcc ctctgaagtt gaagtattgg agtccatcta tctagatgaa
      841 ctacaggtga ttaaaggaaa tggcagaact tcaccatggg agatctacat cactttgcat
      901 cctgccactg cagaggacca ggattcacag tatgtctgct tcactctggt gcttcaggtc
      961 ccagcagagt atccccatga ggtgccacag atctctatcc gaaatccccg aggactttca
     1021 gatgaacaga tccacacgat cttacaggtg ctgggccacg tggccaaggc tgggctgggc
     1081 actgccatgc tgtatgaact cattgagaaa gggaaggaaa ttctcacaga taacaacatc
     1141 cctcatggcc agtgtgtcat ctgcctctat ggtttccagg agaaggaggc ctttaccaaa
     1201 acaccctgtt accactactt ccactgccac tgccttgctc ggtacatcca gcacatggag
     1261 caagagctga aggcacaagg acaggagcag gaacaggaac ggcagcatgc tacaaccaaa
     1321 cagaaggcag tcggtgtgca gtgtccagtg tgcagagagc ccctcgtgta tgatcttgcc
     1381 tcactgaaag cagcccctga accccaacag cccatggagc tgtaccagcc cagtgcagag
     1441 agcttgcgcc agcaagaaga acgcaagcgg ctctaccaga ggcagcagga gcggggggga
     1501 atcattgacc ttgaggctga gcgaaaccga tacttcatca gccttcagca gcctcctgcc
     1561 cctgcggaac ctgagtcagc tgtagatgtc tccaaaggat cccaaccacc cagcaccctt
     1621 gcagcagaac tatccacctc accagccgtc caatccactt tgccacctcc tctgcctgtg
     1681 gcgacccagc acatatgtga gaagattcca gggaccaggt caaatcagca aaggttgggc
     1741 gaaacccaga aagctatgct agatcccccc aagcccagtc gaggtccctg gcgacagccc
     1801 gaacggaggc acccaaaggg aggggagtgc cacgccccta aaggtacccg tgacacccag
     1861 gaactgccac ctcctgaggg gcccctcaag gagcccatgg acctaaagcc agaaccccat
     1921 agccaaggag ttgaaggtcc tccacaagag aaggggcctg gcagctggca ggggccccca
     1981 ccccgcagga ctcgggactg tgttcgctgg gagcgctcta aaggccggac acccggttct
     2041 tcctaccctc gcctgcctcg gggccagtag ctcgagtcta gagggcccgt ttaaacccgc
     2101 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2161 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2221 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2281 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2341 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2401 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2461 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2521 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2581 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2641 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2701 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2761 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2821 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2881 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3001 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3061 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3121 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3181 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3241 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3301 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3361 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3421 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3481 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3541 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3601 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3661 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3721 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3781 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3841 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3901 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3961 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4021 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4081 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4141 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4321 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4381 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4441 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4501 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4561 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4621 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4681 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4741 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4801 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4861 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4921 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4981 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5041 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5101 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5161 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5221 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5281 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5341 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5401 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5461 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5521 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5581 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5641 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5701 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5761 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5821 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5881 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5941 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6001 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6061 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6121 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6181 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6241 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6301 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6361 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6421 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6481 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6541 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6601 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6661 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6721 cttcgcgatg tacgggccag atatacgcgt t
//
