LOCUS       sequence-133140-        7187 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 7187)
  AUTHORS   Ohta T, Xiong Y
  TITLE     Phosphorylation- and Skp1-independent in vitro ubiquitination of 
            E2F1 by multiple ROC-cullin ligases.
  JOURNAL   Cancer Res. 2001 Feb 15. 61(4):1347-53.
  PUBMED    11245432
REFERENCE   2  (bases 1 to 7187)
  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: "Cancer Res.
            2001 Feb 15. 61(4):1347-53."
COMMENT     Sequence Label: pcDNA3-myc-Skp2
FEATURES             Location/Qualifiers
     source          1..7187
                     /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..2863
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4247..4434
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4302..4326)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(4677..4695)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4678..4695)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4715..4732)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4721..4945
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4991..5419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5078..5097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5288..5309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5428..5448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5433..5762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5613..5748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5675..5694
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5829..6623
                     /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(5883..5902)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6493..6512
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6797..6918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6834..6853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6888..6907
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6967..6983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6967..6983)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6980..7002)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6991..7007
                     /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(7015..7021,7022..7039,7040..7045))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7060..7081
                     /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 atggagcaaa agctcatctc agaggaagat ctggaattcc cggggatccc
     2881 catggacgta tttaaaactc ccgggcctgc ggacgctatg cacaggaagc acctccagga
     2941 gattccagac ctgagtagca acgttgccac cagcttcacg tggggatggg attccagcaa
     3001 gacttctgaa ctgctgtcag gcatgggggt ctccgccctg gagaaagagg agcccgacag
     3061 tgagaacatc ccccaggaac tgctctcaaa cctgggccac ccggagagcc ccccacggaa
     3121 acggctgaag agcaaaggga gtgacaaaga ctttgtgatt gtccgcaggc ctaagctaaa
     3181 tcgagagaac tttccaggtg tttcatggga ctcccttccg gatgagctgc tcttgggaat
     3241 cttttcctgt ctgtgcctcc ctgagctgct aaaggtctct ggtgtttgta agaggtggta
     3301 tcgcctagcg tctgatgagt ctctatggca gaccttagac ctcacaggta aaaatctgca
     3361 cccggatgtg actggtcggt tgctgtctca aggggtgatt gccttccgct gcccacgatc
     3421 atttatggac caaccattgg ctgaacattt cagccctttt cgtgtacagc acatggacct
     3481 atcgaactca gttatagaag tgtccaccct ccacggcata ctgtctcagt gttccaagtt
     3541 gcagaatcta agcctggaag gcctgcggct ttcggatccc attgtcaata ctctcgcaaa
     3601 aaactcaaat ttagtgcgac ttaacctttc tgggtgttct ggattctctg aatttgccct
     3661 gcagactttg ctaagcagct gttccagact ggatgagctg aacctctcct ggtgttttga
     3721 tttcactgaa aagcatgtac aggtggctgt tgcgcatgtg tcagagacca tcacccagct
     3781 gaatcttagc ggctacagaa agaatctcca gaaatcagat ctctctactt tagttagaag
     3841 atgccccaat cttgtccatc tagacttaag tgatagtgtc atgctaaaga atgactgctt
     3901 tcaggaaatt ttccagctca actacctcca acacctatca ctcagtcggt gctatgatat
     3961 aatacctgaa actttacttg aacttggaga aattcccaca ctaaaaacac tacaagtttt
     4021 tggaatcgtg ccagatggta cccttcaact gttaaaggaa gcccttcctc atctacagat
     4081 taattgctcc catttcacca ccattgccag gccaactatt ggcaacaaaa agaaccagga
     4141 gatatggggc atcaaatgcc gactgacact gcaaaagccc agttgtctat gaagtattta
     4201 ttgcaggatg gtgtctcgac gagctctgca gccaagctaa ttccgggcga atttcttatg
     4261 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     4321 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     4381 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc
     4441 ggcaagtgca caaacaatac ttaaataaat actactcagt aataacctat ttcttagcat
     4501 ttttgacgaa atttgctatt ttgttagagt cttttacacc atttgtctcc acacctccgc
     4561 ttacatcaac accaataacg ccatttaatc taagcgcatc accaacattt tctggcgtca
     4621 gtccaccagc taacataaaa tgtaagcttc tcgagcatgc atctagaggg ccctattcta
     4681 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     4741 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4801 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4861 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4921 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     4981 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5041 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5281 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5341 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5401 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5461 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5581 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5641 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5701 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5761 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     5821 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5881 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5941 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6001 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6061 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6121 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6181 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6241 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6301 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6361 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6421 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6481 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6541 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6601 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     6661 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6721 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6781 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6901 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
