LOCUS       sequence-258279-        7854 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 7854)
  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 7854)
  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-CUL1
FEATURES             Location/Qualifiers
     source          1..7854
                     /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(3173..3191)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3174..3191)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3211..3228)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3217..3441
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3487..3915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3574..3593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3784..3805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3924..3944)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3929..4258
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4109..4244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4171..4190
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4325..5119
                     /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(4379..4398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4989..5008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5293..5414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5330..5349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5384..5403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5463..5479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5463..5479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5476..5498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5487..5503
                     /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(5511..5517,5518..5535,5536..5541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5556..5577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5694..5711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5865..6453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5945..5964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6624..7415,7416..7484))
                     /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     7247..7266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7485..7589)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7664..7683)
                     /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 tggcgtcaac ccggagccag aacccccacg gcctgaagca gattggcctg gaccagatct
      841 gggacgacct cagagccggc atccagcagg tgtacacacg gcagagcatg gccaagtcca
      901 gatatatgga gctctacact catgtttata actactgtac tagtgttcac cagtcaaacc
      961 aagcacgagg agctggagtt cctccttcta agtcgaaaaa ggggcagaca cctggaggag
     1021 ctcagtttgt tggcctggaa ttatataaac gacttaagga atttttgaag aattacttga
     1081 caaatcttct taaggatgga gaagatttga tggatgagag tgtactgaaa ttctacactc
     1141 aacaatggga agattatcga ttttcaagca aagtgctgaa tggaatttgt gcctacctca
     1201 atagacattg ggttcgccgt gaatgtgacg aaggacgaaa aggaatatat gaaatctatt
     1261 cgcttgcatt ggtgacttgg agagactgtc tgttcaggcc actgaataaa caggtaacaa
     1321 atgctgtttt aaagctgatt gaaaaggaaa ggaatggtga aaccatcaat acaagattga
     1381 ttagtggagt tgtacagtct tacgtggaat tggggctgaa tgaagatgat gcatttgcaa
     1441 agggccctac gttaacagtg tataaagaat cctttgaatc tcaatttttg gctgacacag
     1501 agagatttta taccagagag agtactgaat tcttgcagca gaacccagtt actgaatata
     1561 tgaaaaaggc agaggctcgt ctgcttgagg aacaacgaag agttcaggtt taccttcatg
     1621 aaagcacaca agatgaatta gcaaggaaat gtgaacaagt cctcattgaa aaacacttgg
     1681 aaattttcca cacagaattt cagaatttat tggatgctga caaaaatgaa gatttgggac
     1741 gcatgtataa tcttgtatct agaatccagg atggcctagg agaattgaaa aaactgttgg
     1801 agacacacat tcataatcag ggtcttgcag ccattgaaaa gtgtggagaa gctgctttaa
     1861 atgaccccaa aatgtatgta cagacagtgc ttgatgttca taaaaaatac aatgccctgg
     1921 taatgtctgc attcaacaat gacgctggct ttgtggctgc tcttgataag gcttgtggtc
     1981 gcttcataaa caacaacgcg gttaccaaga tggcccaatc atccagtaaa tcccctgagt
     2041 tgctggctcg atactgtgac tccttgttga agaaaagttc caagaaccca gaggaggcag
     2101 aactagaaga cacactcaat caagtgatgg ttgtcttcaa gtacatagaa gacaaagacg
     2161 tatttcagaa gttctatgcg aagatgctcg ccaagaggct cgtccaccag aacagtgcaa
     2221 gtgacgatgc cgaagccagc atgatctcca agttaaagca agcttgcggg ttcgagtaca
     2281 cctctaaact tcagcgcatg tttcaagaca ttggcgtgag caaagatctg aacgagcaat
     2341 tcaaaaagca cttgacaaac tcagaacccc tagacttgga tttcagcatt caagtgctga
     2401 gctccgggtc ctggcccttc cagcagtctt gtacatttgc cttgccgtca gagttggaac
     2461 gtagttatca gcgattcaca gctttctacg ccagccgcca cagtggccga aaattgacgt
     2521 ggttatatca gttgtctaaa ggagaattgg taactaactg cttcaaaaac agatatactt
     2581 tgcaggcgtc gacattccag atggctatcc tgcttcagta caacacggaa gatgcctaca
     2641 ctgtgcagca gctgaccgac agcactcaaa ttaaaatgga cattttggcg caagttttac
     2701 agattttatt aaagtcgaag ctattggtct tggaagatga aaatgcaaat gttgatgagg
     2761 tggaattgaa gccagatacc ttaataaaat tatatcttgg ttataaaaat aagaaattaa
     2821 gggttaacat caatgtgcca atggaaaccg aacagaagca ggaacaagaa accacacaca
     2881 aaaacatcga ggaagaccgc aaactactga ttcaggcggc catcgtgaga atcatgaaga
     2941 tgaggaaggt tctgaaacac cagcagttac ttggcgaggt cctcactcag ctgtcctcca
     3001 ggttcaaacc tcgagtccct gtgatcaaga aatgcattga cattctaatt gagaaagaat
     3061 atttggagcg agtggatggt gaaaaggaca cctacagtta cttggcttaa cccttctgga
     3121 tcctgatatc cgcggtacca agcttctcga gcatgcatct agagggccct attctatagt
     3181 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     3241 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3301 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3361 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3421 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3481 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3541 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3601 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3661 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3721 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3781 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3841 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3901 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4021 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4081 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4141 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4201 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4261 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4321 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4381 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4441 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4501 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4561 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4621 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4681 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4741 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4801 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4861 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4921 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4981 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5041 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5101 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5161 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5221 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5281 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5401 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5521 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5701 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5761 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5821 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5881 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5941 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6001 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6061 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6301 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7621 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     7681 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7741 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7801 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
