LOCUS       sequence-138779-        8209 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 8209)
  AUTHORS   Hu J, Zacharek S, He YJ, Lee H, Shumway S, Duronio RJ, Xiong Y
  TITLE     WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 
            by DDB1-CUL4-ROC1 ligase.
  JOURNAL   Genes Dev. 2008 Apr 1. 22(7):866-71.
  PUBMED    18381890
REFERENCE   2  (bases 1 to 8209)
  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: "Genes Dev. 
            2008 Apr 1. 22(7):866-71."
COMMENT     Sequence Label: pcDNA3-HA2-CUL4A
FEATURES             Location/Qualifiers
     source          1..8209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2834..2860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2864..2890
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2870..2896
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(5699..5717)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5700..5717)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    5743..5967
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6013..6441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6100..6119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6310..6331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6450..6470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6455..6784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6635..6770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6697..6716
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6851..7645
                     /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(6905..6924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7515..7534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7819..7940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7856..7875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7910..7929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7989..8005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7989..8005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8002..8024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8013..8029
                     /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(8037..8043,8044..8061,8062..8067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8082..8103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct atgtacccat acgatgttcc tgactatgcg ggctatccct atgacgtccc
     2881 ggactatgca agcttggtac cgttaacatg gatcccccgg gctgcaggaa ttcggcacga
     2941 gccagccatg gcggacgagg ccccgcggaa gggcagcttc tcggcgctcg tgggccgcac
     3001 caacggcctc accaagcccg cggccctggc cgccgcgccc gccaagccgg ggggcgcggg
     3061 cggctccaag aagctggtca tcaagaactt ccgagacaga cctcggctgc ccgacaacta
     3121 cacgcaggac acgtggcgga agctgcacga ggcggtgcgg gccgtgcaga gcagcacctc
     3181 catcaggtac aacctcgagg agctctacca ggctgtggaa aatctctgtt ctcacaaagt
     3241 ctccccaatg ctctacaagc aactgcgtca ggcctgtgaa gaccacgtcc aggcacagat
     3301 ccttccgttt agagaagact cactagatag tgttttattt ttaaagaaga ttaacacgtg
     3361 ctggcaggac cactgcagac aaatgatcat gatcagaagc atcttcctgt tcttggaccg
     3421 cacctatgtg ctgcagaact ccacgctgcc ctccatctgg gatatgggat tagaactgtt
     3481 tagaacccat attattagtg ataaaatggt tcagagtaaa accattgatg gaatcctact
     3541 gctgatcgag cgcgagagga gcggcgaggc cgtggaccgg agcctgttgc ggagcctcct
     3601 gggcatgctg tctgacctgc aggtgtataa agattcattt gaactgaaat ttttggaaga
     3661 gactaattgc ttatatgctg ccgaaggcca aaggttaatg caggaaagag aggtcccaga
     3721 atatcttaac catgtaagta aacgcttaga ggaagaggga gacagagtaa tcacttactt
     3781 ggaccacagc acacagaaac cactgattgc ttgtgtggag aaacagctat taggagaaca
     3841 tttaacagca attctgcaga aagggctcga ccacttactg gatgagaaca gagtgccgga
     3901 cctcgcacag atgtaccagc tgttcagccg ggtgaggggc gggcagcagg cgctgctgca
     3961 gcactggagc gagtacatca agacttttgg aacagcgatc gtaatcaatc ctgagaaaga
     4021 taaagacatg gtccaagacc tgttggactt caaggacaag gtggaccacg tgatcgaggt
     4081 ctgcttccag aagaatgagc ggttcgtcaa cctgatgaag gagtcctttg agacgttcat
     4141 caacaagaga cccaacaagc ctgcagaact gatcgcaaag catgtggatt caaagttaag
     4201 agcaggcaac aaagaagcca cagacgagga gctggagcgg acgttggaca agatcatgat
     4261 cctgttcagg tttatccacg gtaaagatgt ctttgaagca ttttataaaa aagatttggc
     4321 aaaaagactc cttgttggga aaagtgcctc agtcgatgct gaaaagtcta tgttgtcaaa
     4381 gctcaagcat gagtgcggtg cagccttcac cagcaagctg gaaggcatgt tcaaggacat
     4441 ggagctttcg aaggacatca tggttcattt caagcagcat atgcagaatc agagtgactc
     4501 aggccctata gacctcacag tgaacatact cacaatgggc tactggccaa catacacgcc
     4561 catggaagtg cacttaaccc cagaaatgat taaacttcag gaagtattta aggcatttta
     4621 tcttggaaag cacagtggtc gaaaacttca gtggcaaact actttgggac atgctgtttt
     4681 aaaagcggag tttaaagaag ggaagaagga attccaggtg tccctcttcc agacactggt
     4741 gctcctcatg ttcaacgagg gagatggctt cagctttgag gagataaaaa tggccacggg
     4801 gatagaggat agtgaattgc gcagaacgct gcagtccctg gcctgtggca aagcacgtgt
     4861 gctgattaaa agtcccagag gaaaggaagt ggaagatgga gacaagttca tttttaatgg
     4921 agagttcaag cacaagttgt ttagaataaa gatcaatcaa attcagatga aggaaactgt
     4981 tgaggaacag gttagcacca ctgagagagt gtttcaggat agacaatatc agattgatgc
     5041 tgctatcgtc agaataatga agatgagaaa gactcttggt cataatcttc tagtttctga
     5101 attatataat cagctgaaat ttccagtaaa gcctggagat ttgaaaaaga gaattgaatc
     5161 tctgatagac agagactata tggagagaga caaagacaat ccgaatcagt accactacgt
     5221 ggcctgacgc atctgcagac ggttcccctt catgaaacac tagaatgtac cctcagagca
     5281 ggaagcacac ctgtgccatt tctgggactc tgattgatcc agctgtggac attggaaggc
     5341 gaaggaaggg aggtggctcc tgggtcatct ttcacaaggc tcaagacttc aacctgcaga
     5401 tgtatctttt tccctccagt ttttcctcta gttcttttag gcatttaaat tgtttctgtt
     5461 actctgtgca aaataacttt gagattggac aagaagatgt tactaaagag aagttccttt
     5521 aaaaggtctt gttcttgtgt caaaaagctg caagtttggt ttgttctcgt gtgtgatcat
     5581 gagtgcacaa tgaagaagac cctagatgct gcatttttta gctctgaaga ttccttaggt
     5641 atccctgaag acagctcgct cagatgatca gcatttagag tgaaaacaag ggccctattc
     5701 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     5761 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5821 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5881 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5941 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     6001 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6061 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6181 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6301 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6361 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6421 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6721 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6781 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     6841 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     6901 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6961 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     7021 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     7081 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     7141 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     7201 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     7261 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     7321 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     7381 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     7441 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     7501 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     7561 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     7621 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     7681 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7741 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7801 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7921 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     8101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//
