LOCUS       sequence-169081-        7408 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 7408)
  AUTHORS   van den Heuvel S, Harlow E
  TITLE     Distinct roles for cyclin-dependent kinases in cell cycle control.
  JOURNAL   Science 1993 Dec 24;262(5142):2050-4.
  PUBMED    8266103
REFERENCE   2  (bases 1 to 7408)
  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: "Science"; 
            date: "1993-12-24"; volume: "262(5142)"; pages: "2050-4"
COMMENT     Sequence Label: Cdk5-HA
FEATURES             Location/Qualifiers
     source          1..7408
                     /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     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2330..2349)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2395..2450
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2449..2468)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2808..2824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2808..2824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2821..2843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2832..2848
                     /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(2856..2862,2863..2880,2881..2886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2901..2922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3039..3056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3210..3798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3290..3309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3969..4760,4761..4829))
                     /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     4592..4611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4830..4934)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5002..5020
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5058..5080)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5180..5199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5393..5415
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5407..5424
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5408..5424
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5835..5980
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6037..6831
                     /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(6091..6110)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6701..6720
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7025..7044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7069..7116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atccatgcag aaatacaaga aactggaaaa gattggggaa ggcacctacg
     1381 gaactgtgtt caaggccaaa aaccgggaga ctcatgagat cgtggctctg aaacgggtga
     1441 ggctggatga cgatgatgag ggtgtgccga gttccgccct ccgggagatc tgcctactca
     1501 aggagctgaa gcacaagaac atcgtcaggc ttcatgacgt cctgcacagc gacaagaagc
     1561 tgactttggt ttttgaattc tgtgaccagg acctgaagaa gtattttgac agttgcaatg
     1621 gtgacctcga tcctgagatt gtaaagtcat tcctcttcca gctactaaaa gggctgggat
     1681 tctgtcatag ccgcaatgtg ctacacaggg acctgaagcc ccagaacctg ctaataaaca
     1741 ggaatgggga gctgaaattg gctgattttg gcctggctcg agcctttggg attcccgtcc
     1801 gctgttactc agctgaggtg gtcacactgt ggtaccgccc accggatgtc ctctttgggg
     1861 ccaagctgta ctccacgtcc atcgacatgt ggtcagccgg ctgcatcttt gcagagctgg
     1921 ccaatgttgg gcggcctctt tttcccggca atgatgtcga tgaccagttg aagaggatct
     1981 tccgactgct ggggacgccc accgaggagc agtggccctc tatgaccaag ctgccagact
     2041 ataagcccta tccgatgtac ccggccacaa catccctggt gaacgtcgtg cccaaactca
     2101 atgccacagg gagggatctg ctgcagaacc ttctgaagtg taaccctgtc cagcgtatct
     2161 cagcagaaga ggccctgcag cacccctact tctccgactt ctgtccgccc tatgatgtgc
     2221 cagattatgc ttagggatcc gtcgaggaat tcactcctca ggtgcaggct gcctatcaga
     2281 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     2341 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     2401 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     2461 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     2521 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     2581 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     2641 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     2701 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     2761 ttctcttatg gagatccctc gacggatcgg ccgcaattcg taatcatgtc atagctgttt
     2821 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2881 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2941 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3001 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3061 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3121 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3181 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3241 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3301 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3361 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3421 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3481 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3541 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3601 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3661 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3721 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3781 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3841 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3901 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3961 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4021 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4081 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4141 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4201 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4261 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4321 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4381 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4441 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4501 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4561 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4621 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4681 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4741 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4801 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4861 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4921 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4981 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5041 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5101 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5161 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5221 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5281 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5341 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5401 cgacgttgta aaacgacggc cagtgccaag cttcacgctg ccgcaagcac tcagggcgca
     5461 agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc
     5521 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     5581 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     5641 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     5701 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tcctgcttca
     5761 tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg
     5821 tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca
     5881 cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg
     5941 tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc
     6001 agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6061 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6121 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6181 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6241 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6301 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6361 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6421 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6481 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6541 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6601 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6661 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6721 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6781 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6841 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6961 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggaga tgggggaggc
     7021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     7081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     7141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     7201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     7261 gggcggcaag ccctgccata gccacgggcc ccgtgggtta gggacggcgg atcgcggccc
     7321 tctagagagc ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt
     7381 ggctcatgtc caacattacc gccatgtt
//
