LOCUS       sequence-426783-        7604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-CDT2 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7604)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 7604)
  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. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: FLAG-CDT2
FEATURES             Location/Qualifiers
     source          1..7604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             926..949
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3195..3212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3201..3425
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3471..3899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3768..3789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3908..3928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3913..4242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4093..4228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4155..4174
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4309..5103
                     /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(4363..4382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4973..4992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5277..5398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5314..5333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5368..5387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5447..5463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5447..5463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5460..5482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5471..5487
                     /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(5495..5501,5502..5519,5520..5525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5540..5561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5678..5695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5849..6437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5929..5948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6608..7399,7400..7468))
                     /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     7231..7250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7469..7573)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggatta caaggatgac gacgataagg gtaccgagct
      961 cggatccctc ttcaattcgg tgctccgcca gccccagctt ggcgtcctga gaaatggatg
     1021 gtcttcacaa taccctcttc aatcccttct gactggttat cagtgcagtg gtaatgatga
     1081 acacacttct tatggagaaa caggagtccc agttcctcct tttggatgta ccttctcttc
     1141 tgctcccaat atggaacatg tactagcagt tgccaatgaa gaaggctttg ttcgattgta
     1201 taacacagaa tcacaaagtt tcagaaagaa gtgcttcaaa gaatggatgg ctcactggaa
     1261 tgccgtcttt gacctggcct gggttcctgg tgaacttaaa cttgttacag cagcaggtga
     1321 tcaaacagcc aaattttggg acgtaaaagc tggtgagctg attggaacat gcaaaggtca
     1381 tcaatgcagc ctcaagtcag ttgccttttc taagtttgag aaagctgtat tctgtacggg
     1441 tggaagagat ggcaacatta tggtctggga taccaggtgc aacaaaaaag atgggtttta
     1501 taggcaagtg aatcaaatca gtggagctca caatacctca gacaagcaaa ccccttcaaa
     1561 acccaagaag aaacagaatt caaaaggact tgctccttct gtggatttcc agcaaagtgt
     1621 tactgtggtc ctctttcaag acgagaatac cttagtctca gcaggagctg tggatgggat
     1681 aatcaaagta tgggatttac gtaagaatta tactgcttat cgacaagaac ccatagcatc
     1741 caagtctttc ctgtacccag gtagcagcac tcgaaaactt ggatattcaa gtctgatttt
     1801 ggattccact ggctctactt tatttgctaa ttgcacagac gataacatct acatgtttaa
     1861 tatgactggg ttgaagactt ctccagtggc tattttcaat ggacaccaga actctacctt
     1921 ttatgtaaaa tccagcctta gtccagatga ccagttttta gtcagtggct caagtgatga
     1981 agctgcctac atatggaagg tctccacacc ctggcaacct cctactgtgc tcctgggtca
     2041 ttctcaagag gtcacgtctg tgtgctggtg tccatctgac ttcacaaaga ttgctacctg
     2101 ttctgatgac aatacactaa aaatctggcg cttgaataga ggcttagagg agaaaccagg
     2161 aggtgataaa ctttccacgg tgggttgggc ctctcagaag aaaaaagagt caagacctgg
     2221 cctagtaaca gtaacgagta gccagagtac tcctgccaaa gcccccaggg caaagtgcaa
     2281 tccatccaat tcttccccgt catccgcagc ttgtgcccca agctgtgctg gagacctccc
     2341 tcttccttca aatactccta cgttctctat taaaacctct cctgccaagg cccggtctcc
     2401 catcaacaga agaggctctg tctcctccgt ctctcccaag ccaccttcat ctttcaagat
     2461 gtcgattaga aactgggtga cccgaacacc ttcctcatca ccacccatca ctccacctgc
     2521 ttcggagacc aagatcatgt ctccgagaaa agcccttatt cctgtgagcc agaagtcatc
     2581 ccaagcagag gcttgctctg agtctagaaa tagagtaaag aggaggctag actcaagctg
     2641 tctggagagt gtgaaacaaa agtgtgtgaa gagttgtaac tgtgtgactg agcttgatgg
     2701 ccaagttgaa aatcttcatt tggatctgtg ctgccttgct ggtaaccagg aagaccttag
     2761 taaggactct ctaggtccta ccaaatcaag caaaattgaa ggagctggta ccagtatctc
     2821 agagcctccg tctcctatca gtccgtatgc ttcagaaagc tgtggaacgc tacctcttcc
     2881 tttgagacct tgtggagaag ggtctgaaat ggtaggcaaa gagaatagtt ccccagagaa
     2941 taaaaactgg ttgttggcca tggcagccaa acggaaggct gagaatccat ctccacgaag
     3001 tccgtcatcc cagacaccca attccaggag acagagcgga aagaaattgc caagcccggt
     3061 caccatcacg cccagctcca tgaggaaaat ctgcacatac ttccatagaa agtcccagga
     3121 ggacttctgt ggtcctgaac actcaacaga attatagctc gagtctagag ggcccgttta
     3181 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3241 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3301 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3361 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3421 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3901 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4261 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4321 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4381 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4441 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4501 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4561 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4621 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4681 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4741 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4801 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4861 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4921 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4981 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5041 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5101 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5161 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5221 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5281 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5341 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5401 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5461 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5521 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5581 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5641 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5701 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5761 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5821 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5881 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5941 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6001 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6061 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6541 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6601 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6661 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6721 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6781 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6841 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6901 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6961 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7021 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7081 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7141 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7201 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7261 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7321 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7381 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7441 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7501 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7561 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
