LOCUS       sequence-288015-        7510 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 7510)
  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 7510)
  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: Cdk6-HA
FEATURES             Location/Qualifiers
     source          1..7510
                     /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(2432..2451)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2497..2552
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2551..2570)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2910..2926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2910..2926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2923..2945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2934..2950
                     /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(2958..2964,2965..2982,2983..2988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3003..3024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3141..3158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3312..3900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3392..3411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4071..4862,4863..4931))
                     /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     4694..4713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4932..5036)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5104..5122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5160..5182)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5282..5301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5495..5517
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5509..5526
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5510..5526
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5937..6082
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6139..6933
                     /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(6193..6212)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6803..6822
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7127..7146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7171..7218
                     /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 atccatggag aaggacggcc tgtgccgcgc tgaccagcag tacgaatgcg
     1381 tggcggagat cggggagggc gcctatggga aggtgttcaa ggcccgcgac ttgaagaacg
     1441 gaggccgttt cgtggcgttg aagcgcgtgc gggtgcagac cggcgaggag ggcatgccgc
     1501 tctccaccat ccgcgaggtg gcggtgctga ggcacctgga gaccttcgag caccccaacg
     1561 tggtcaggtt gtttgatgtg tgcacagtgt cacgaacaga cagagaaacc aaactaactt
     1621 tagtgtttga acatgtcgat caagacttga ccacttactt ggataaagtt ccagagcctg
     1681 gagtgcccac tgaaaccata aaggatatga tgtttcagct tctccgaggt ctggactttc
     1741 ttcattcaca ccgagtagtg catcgcgatc taaaaccaca gaacattctg gtgaccagca
     1801 gcggacaaat aaaactcgct gacttcggcc ttgcccgcat ctatagtttc cagatggctc
     1861 taacctcagt ggtcgtcacg ctgtggtaca gagcacccga agtcttgctc cagtccagct
     1921 acgccacccc cgtggatctc tggagtgttg gctgcatatt tgcagaaatg tttcgtagaa
     1981 agcctctttt tcgtggaagt tcatatgttg atcaactagg aaaaatcttg gacgtgattg
     2041 gactcccagg agaagaagac tggcctagag atgttgccct tcccaggcag gcttttcatt
     2101 caaaatctgc ccaaccaatt gagaagtttg taacagatat cgatgaacta ggcaaagacc
     2161 tacttctgaa gtgtttgaca tttaacccag ccaaaagaat atctgcctac agtgccctgt
     2221 ctcacccata cttccaggac ctggaaaggt gcaaagaaaa cctggattcc cacctgccgc
     2281 ccagccagaa cacctcggag ctgaatacag cctatgatgt gccagattat gcttgaggat
     2341 ccgtcgagga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     2401 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     2461 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     2521 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2581 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     2641 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     2701 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     2761 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     2821 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     2881 tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     2941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3001 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3061 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3121 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3181 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3241 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3301 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3721 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3901 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3961 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4201 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4381 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4681 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5041 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5101 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     5161 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5221 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     5281 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     5341 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     5401 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     5461 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     5521 gccagtgcca agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     5581 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     5641 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     5701 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     5761 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     5821 cgccaaggat ctgatggcgc aggggatcaa gatcctgctt catccccgtg gcccgttgct
     5881 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
     5941 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
     6001 ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
     6061 gaccctgcag cgacccgctt aacagcgtca acagcgtgcc gcagatctga tcaagagaca
     6121 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6181 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6241 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6301 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6361 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6421 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6481 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6541 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6601 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6661 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6721 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6781 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6841 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6901 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6961 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7021 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7081 tcatgctgga gttcttcgcc caccccggga gatgggggag gctaactgaa acacggaagg
     7141 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     7201 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     7261 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     7321 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca agccctgcca
     7381 tagccacggg ccccgtgggt tagggacggc ggatcgcggc cctctagaga gcttggccca
     7441 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     7501 ccgccatgtt
//
