LOCUS       sequence-139465-        7733 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 7733)
  AUTHORS   Ohta T, Michel JJ, Schottelius AJ, Xiong Y
  TITLE     ROC1, a homolog of APC11, represents a family of cullin partners 
            with an associated ubiquitin ligase activity.
  JOURNAL   Mol Cell. 1999 Apr . 3(4):535-41.
  PUBMED    10230407
REFERENCE   2  (bases 1 to 7733)
  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: "Mol Cell. 
            1999 Apr . 3(4):535-41."
COMMENT     Sequence Label: pcDNA3-myc3-CUL2
FEATURES             Location/Qualifiers
     source          1..7733
                     /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             663..692
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             699..728
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             735..764
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3052..3070)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3053..3070)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3090..3107)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3096..3320
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3366..3794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3453..3472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3663..3684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3803..3823)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3808..4137
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3988..4123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4050..4069
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4204..4998
                     /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(4258..4277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4868..4887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5172..5293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5209..5228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5263..5282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5342..5358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5342..5358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5355..5377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5366..5382
                     /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(5390..5396,5397..5414,5415..5420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5435..5456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5573..5590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5744..6332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5824..5843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6503..7294,7295..7363))
                     /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     7126..7145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7364..7468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7543..7562)
                     /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 acccaagcta
      661 tggagcaaaa gctcatctca gaggaagatc tggaattaga gcaaaagctc atctcagagg
      721 aagatctgga attagagcaa aagctcatct cagaggaaga tctggaattc aggatcccca
      781 tgtctttgaa accaagagta gtagattttg atgaaacatg gaacaaactt ttgacgacaa
      841 taaaagccgt ggtcatgttg gaatacgtcg aaagagcaac atggaatgac cgtttctcag
      901 atatctatgc tttatgtgtg gcctatcctg aaccccttgg agaaagactt tatacagaaa
      961 ctaagatttt tttggaaaat catgttcggc atttgcataa gagagttttg gagtcagaag
     1021 aacaagtact tgttatgtat cataggtact gggaagaata cagcaagggt gcagactata
     1081 tggactgctt atataggtat ctcaacaccc agtttattaa aaagaataaa ttaacagaag
     1141 cggaccttca gtatggctat ggtggtgtag atatgaatga accacttatg gaaataggag
     1201 agctagcatt ggatatgtgg aggaaattga tggttgaacc acttcaggcc atccttatcc
     1261 gaatgctgct ccgagaaatc aaaaatgatc gtggtggaga agacccaaac cagaaagtaa
     1321 tccatggggt tattaactcc tttgttcatg ttgaacagta taagaaaaaa ttccccttaa
     1381 agttttatca ggaaattttt gagtctccct ttctgactga aacaggagag tattacaaac
     1441 aagaagcttc aaatttatta caagaatcaa actgctcaca gtatatggaa aaggttctag
     1501 gtagattaaa agatgaagaa attcgatgtc gaaaatacct acatccaagt tcgtatacta
     1561 aggtgattca tgaatgtcaa caacgaatgg tagcagacca cttacagttt ttacatgcag
     1621 aatgtcataa tataattcga caagagaaaa aaaatgacat ggcaaatatg tacgtcttac
     1681 tccgtgctgt gtccactggt ttacctcata tgattcagga gctgcaaaac cacatccatg
     1741 atgagggcct tcgagcaacc agcaacctta ctcaggaaaa catgccaaca ctatttgtgg
     1801 agtcagtttt ggaagtgcat ggtaaatttg ttcagcttat caacactgtt ttgaatggtg
     1861 atcagcattt tatgagtgcg ttggataagg cccttacgtc agttgtaaat tacagagaac
     1921 ctaagtctgt ttgcaaagca cctgaactgc ttgctaagta ctgtgacaac ttactgaaga
     1981 agtcagcgaa agggatgaca gagaatgaag tggaagacag gctcacgagc ttcatcacag
     2041 tgttcaaata cattgatgac aaggacgtct ttcaaaagtt ctacgcaaga atgctggcaa
     2101 aacgtttaat tcatgggtta tccatgtcta tggactctga agaagccatg atcaacaaat
     2161 taaagcaagc ctgtggttat gagtttacca gcaagctaca tcggatgtat acagatatga
     2221 gtgtcagcgc tgatctcaac aataagttca acaattttat caaaaaccaa gacacagtaa
     2281 tagatttggg aattagtttt caaatatatg ttctacaggc tggtgcgtgg cctcttactc
     2341 aggctccttc atctacgttt gcaattcccc aggaattaga aaaaagtgta cagatgtttg
     2401 aattatttta tagccaacat ttcagtggaa ggaaacttac atggttacat tatctgtgta
     2461 caggtgaagt taaaatgaac tatttgggca aaccatatgt agccatggtt acaacatacc
     2521 aaatggcagt tcttcttgcc tttaacaaca gtgaaactgt cagttataaa gagcttcagg
     2581 acagcactca gatgaatgaa aaggaactga caaaaacaat caaatcatta cttgatgtga
     2641 aaatgattaa ccatgattca gaaaaggaag atattgatgc agaatcttcg ttttcattaa
     2701 atatgaactt tagcagtaaa agaacaaaat ttaaaattac tacatcaatg cagaaagaca
     2761 caccacaaga aatggagcag actagaagtg cagttgatga ggaccggaaa atgtatctcc
     2821 aagctgctat agttcgtatc atgaaagcac gaaaagtgct tcggcacaat gcccttattc
     2881 aagaggtgat tagccagtca agagctaggt ttaatcccag tatcagcatg attaagaagt
     2941 gtattgaagt tctgatagac aaacaataca tagaacgcag ccaggcgtcg gcagatgaat
     3001 acagctacgt cgcgtgatgt cgctctcgag catgcatcta gagggcccta ttctatagtg
     3061 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3121 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3181 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3241 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3301 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3361 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3421 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3481 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3541 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3601 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3661 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3721 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3781 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3841 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4021 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4081 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4141 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4201 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4261 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4321 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4381 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4441 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4501 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4561 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4621 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4681 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4741 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4801 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4861 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4921 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4981 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5041 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5101 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5161 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5221 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5281 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6181 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7501 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7561 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7621 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7681 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
