LOCUS       sequence-225640-        7034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Cdt1 fused to HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  AUTHORS   Tsanov N, Kermi C, Coulombe P, Van der Laan S, Hodroj D, Maiorano D
  TITLE     PIP degron proteins, substrates of CRL4Cdt2, and not PIP boxes, 
            interfere with DNA polymerase eta and kappa focus formation on UV 
            damage.
  JOURNAL   Nucleic Acids Res. 2014 Apr;42(6):3692-706. doi: 
            10.1093/nar/gkt1400. Epub 2014 Jan 14.
  PUBMED    24423875
REFERENCE   2  (bases 1 to 7034)
  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"; doi: 
            "10.1093/nar/gkt1400"; journalName: "Nucleic Acids Res"; date: 
            "2014-04"; volume: "42"; issue: "6"; pages: "3692-706"
COMMENT     Sequence Label: pcDNA3-hCdt1-HA
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             987..1259
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     CDS             2544..2570
                     /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(2589..2607)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2590..2607)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2627..2644)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2633..2857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2903..3331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2990..3009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3200..3221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3340..3360)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3345..3674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3525..3660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3741..4535
                     /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(3795..3814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4405..4424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4709..4830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4746..4765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4800..4819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4879..4895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4879..4895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4892..4914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4903..4919
                     /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(4927..4933,4934..4951,4952..4957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5110..5127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttacca
      901 tggagcagcg ccgcgtcacc gacttcttcg cgcgccgccg ccccgggccc ccccgcatcg
      961 cgccgcccaa gctggcctgc cgcaccccca gccccgccag gcccgcactc cgcgccccgg
     1021 cctccgctac cagtggcagc cgcaagcgcg cccgcccgcc cgccgccccc ggacgcgacc
     1081 aggccaggcc accggcccgc aggagactgc ggctgtcggt ggacgaggtt tccagcccca
     1141 gtacccccga ggccccagac atcccagcct gcccttctcc gggccagaag ataaagaaat
     1201 ccaccccggc agcaggtcag ccgccccacc tgacatccgc gcaggaccag gacaccatct
     1261 ctgagcttgc gtcatgcctg caacgggccc gggagctggg ggcaagagtc cgggcgctga
     1321 aggccagtgc ccaggatgct ggggagtcct gcaccccaga ggccgagggc cgccctgagg
     1381 agccatgtgg cgagaaggcg cccgcctacc agcgcttcca tgccctggcc cagcccggcc
     1441 tgccgggact cgtgctgccc tacaagtacc aggtgctggc ggagatgttc cgcagcatgg
     1501 acaccatcgt gggcatgctc cacaaccgct ccgagacgcc cacctttgcc aaggtccagc
     1561 ggggcgtcca ggacatgatg cgtaggcgtt ttgaggagcg caatgttggc cagatcaaaa
     1621 ccgtgtaccc ggcctcctac cgcttccgcc aggagcgcag tgtccccacc ttcaaggatg
     1681 gcaccaggag gtcagattac cagctcacca tcgagccact gctggagcag gaggctgacg
     1741 gagcagcccc ccagctcacg gcctcgcgcc tcctgcagcg acggcagatc ttcagccaga
     1801 agctggtgga gcacgtcaag gagcaccaca aggccttcct ggcctccctg agccccgcca
     1861 tggtggtgcc ggaggaccag ctgacccgct ggcacccgcg cttcaacgtg gatgaagtac
     1921 ccgacatcga gccggccgcg ctgccccagc cacccgccac ggagaagctc accactgctc
     1981 aggaggtgct ggcccgggcc cgcaacctga tttcacccag gatggagaag gccttgagtc
     2041 aattggccct gcgctctgct gcgcccagca gccccgggtc tcccaggcca gcactgccgg
     2101 ctaccccacc agccaccccg cctgcagcct ctcccagtgc tctgaagggg gtgtcccagg
     2161 atctgctgga gcggatccga gccaaggagg cacagaagca gctggcacag atgacgcggt
     2221 gcccggagca ggagcagcgg ctgcagcgct tagaacggct gcctgagctg gcccgcgtgc
     2281 tgcggagcgt ctttgtgtcc gaacgcaagc ctgcgctcag catggaggtg gcctgtgcca
     2341 ggatggtggg cagctgttgt actatcatga gccctgggga aatggagaag cacctgctgc
     2401 tcctctccga gctgctgccg gactggctca gcctccaccg catccgcacc gacacctacg
     2461 tcaagctgga caaggccgcg gacctcgccc acatcactgc acgcctggcc caccagacac
     2521 gtgctgagga ggggctgaat tcttatgatg ttcctgatta tgctagcctc taatctagag
     2581 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     2641 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2701 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2761 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2821 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2881 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3301 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3481 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3601 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3661 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3901 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4261 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct gacg
//
