LOCUS       sequence-222935-        7451 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 7451)
  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 7451)
  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: Cdc2-HA
FEATURES             Location/Qualifiers
     source          1..7451
                     /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(2373..2392)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2438..2493
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2492..2511)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2851..2867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2851..2867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2864..2886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2875..2891
                     /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(2899..2905,2906..2923,2924..2929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3082..3099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3253..3841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3333..3352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4012..4803,4804..4872))
                     /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     4635..4654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4873..4977)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5045..5063
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5101..5123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5223..5242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5436..5458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5450..5467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5451..5467
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5878..6023
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6080..6874
                     /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(6134..6153)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6744..6763
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7068..7087)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7112..7159
                     /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 atccactatg gaagattata ccaaaataga gaaaattgga gaaggtacct
     1381 atggagttgt gtataagggt agacacaaaa ctacaggtca agtggtagcc atgaaaaaaa
     1441 tcagactaga aagtgaagag gaaggggttc ctagtactgc aattcgggaa atttctctat
     1501 taaaggaact tcgtcatcca aatatagtca gtcttcagga tgtgcttatg caggattcca
     1561 ggttatatct catctttgag tttctttcca tggatctgaa gaaatacttg gattctatcc
     1621 ctcctggtca gtacatggat tcttcacttg ttaagagtta tttataccaa atcctacagg
     1681 ggattgtgtt ttgtcactct agaagagttc ttcacagaga cttaaaacct caaaatctct
     1741 tgattgatga caaaggaaca attaaactgg ctgattttgg ccttgccaga gcttttggaa
     1801 tacctatcag agtatataca catgaggtag taacactctg gtacagatct ccagaagtat
     1861 tgctggggtc agctcgttac tcaactccag ttgacatttg gagtataggc accatatttg
     1921 ctgaactagc aactaagaaa ccacttttcc atggggattc agaaattgat caactcttca
     1981 ggattttcag agctttgggc actcccaata atgaagtgtg gccagaagtg gaatctttac
     2041 aggactataa gaatacattt cccaaatgga aaccaggaag cctagcatcc catgtcaaaa
     2101 acttggatga aaatggcttg gatttgctct cgaaaatgtt aatctatgat ccagccaaac
     2161 gaatttctgg caaaatggca ctgaatcatc catattttaa tgatttggac aatcagatta
     2221 agaagatgta tgatgtgcca gattatgctt agctttctga caaaaagttt ccatagtgga
     2281 tccgtcgagg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2341 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     2401 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     2461 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     2521 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     2581 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     2641 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     2701 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     2761 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     2821 ctcgacggat cggccgcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     2881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4981 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5041 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5101 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5161 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5221 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5281 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5341 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5401 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5461 ggccagtgcc aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     5521 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     5581 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     5641 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     5701 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     5761 ccgccaagga tctgatggcg caggggatca agatcctgct tcatccccgt ggcccgttgc
     5821 tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg
     5881 acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg
     5941 cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag
     6001 cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatctg atcaagagac
     6061 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6121 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6181 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6241 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6301 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     6361 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     6421 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6481 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6541 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6601 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6661 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6721 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6781 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6841 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6901 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6961 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     7021 ctcatgctgg agttcttcgc ccaccccggg agatggggga ggctaactga aacacggaag
     7081 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     7141 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     7201 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     7261 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aagccctgcc
     7321 atagccacgg gccccgtggg ttagggacgg cggatcgcgg ccctctagag agcttggccc
     7381 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
     7441 accgccatgt t
//
