LOCUS       sequence-468706-        6599 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 6599)
  AUTHORS   Brignone C, Bradley KE, Kisselev AF, Grossman SR
  TITLE     A post-ubiquitination role for MDM2 and hHR23A in the p53 
            degradation pathway.
  JOURNAL   Oncogene. 2004 May 20. 23(23):4121-9.
  PUBMED    15064742
REFERENCE   2  (bases 1 to 6599)
  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: "Oncogene. 
            2004 May 20. 23(23):4121-9."
COMMENT     Sequence Label: pcDNA3 V5 hHR23A
FEATURES             Location/Qualifiers
     source          1..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     639..659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..752
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1967..2008
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2018..2035
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2058..2075)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2064..2288
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2334..2762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2421..2440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2631..2652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2771..2791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2776..3105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2956..3091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3018..3037
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3172..3966
                     /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(3226..3245)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3836..3855
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4179..4198)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4233..4252
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4312..4328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4312..4328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4325..4347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4336..4352
                     /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(4360..4366,4367..4384,4385..4390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4405..4426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4543..4560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4714..5302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4794..4813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5473..6264,6265..6333))
                     /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     6096..6115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6334..6438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6513..6532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
       61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      721 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt taagcttggt
      781 accgagctcg gatccagtac ccttcaccat ggccgtcacc atcacgctca aaacgctgca
      841 gcagcagacc ttcaagatcc gcatggagcc tgacgagacg gtgaaggtgc taaaggagaa
      901 gatagaagct gagaagggtc gtgatgcctt ccccgtggct ggacagaaac tcatctatgc
      961 cggcaagatc ttgagtgacg atgtccctat cagggactat cgcatcgatg agaagaactt
     1021 tgtggtcgtc atggtgacca agaccaaagc cggccagggt acctcagcac ccccagaggc
     1081 ctcacccaca gctgccccag agtcctctac atccttcccg cctgccccca cctcaggcat
     1141 gtcccatccc ccacctgccg ccagagagga caagagccca tcagaggaat ccgcccccac
     1201 gacgtcccca gagtctgtgt caggctctgt tccctcttca ggtagcagcg ggcgagagga
     1261 agacgcggcc tccacgctag tgacgggctc tgagtatgag acgatgctga cggagatcat
     1321 gtccatgggc tatgagcgag agcgggtcgt ggccgccctg agagccagct acaacaaccc
     1381 ccaccgagcc gtggagtatc tgctcacggg aattcctggg agccccgagc cggaacacgg
     1441 ttctgtccag gagagccagg tatcggagca gccggccacg gaagcaggag agaaccccct
     1501 ggagttcctg cgggaccagc cccagttcca gaacatgcgg caggtgattc agcagaaccc
     1561 tgcgctgctg cccgccctgc tccagcagct gggccaggag aaccctcagc ttttacagca
     1621 aatcagccgg caccaggagc agttcatcca gatgctgaac gagccccctg gggagctggc
     1681 ggacatctca gatgtggagg gggaggtggg cgccatagga gaggaggccc cgcagatgaa
     1741 ctacatccag gtgacgccgc aggagaaaga agctatagag aagttgaagg ccctgggctt
     1801 cccagagagc ctggtcatcc aggcctattt cgcgtgtgaa aaaaatgaga acttggctgc
     1861 caacttcctc ccgagtcaga actttgatga cgagaagggt caagacaatt ctgcagatat
     1921 ccagcacagt ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     1981 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     2041 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2101 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2161 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2221 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2281 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2401 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2461 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2521 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2641 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2701 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2761 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3121 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3181 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3241 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3301 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3361 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3421 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3481 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3541 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3601 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3661 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3721 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3781 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3841 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3901 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3961 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     4021 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4081 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4141 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4261 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4321 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4381 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4501 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4561 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4621 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5521 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5581 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5641 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5701 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5761 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5821 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5881 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5941 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6001 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6061 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6121 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6181 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6241 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6301 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6361 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6421 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6481 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6541 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaat
//
