LOCUS       sequence-229056-        7658 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 7658)
  AUTHORS   Golsteyn RM, Schultz SJ, Bartek J, Ziemiecki A, Ried T, Nigg EA
  TITLE     Cell cycle analysis and chromosomal localization of human Plk1, a 
            putative homologue of the mitotic kinases Drosophila polo and 
            Saccharomyces cerevisiae Cdc5.
  JOURNAL   J Cell Sci. 1994 Jun;107 ( Pt 6):1509-17.
  PUBMED    7962193
REFERENCE   2  (bases 1 to 7658)
  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: "J Cell 
            Sci."; date: "1994-06"; volume: "107 ( Pt 6)"; pages: "1509-17"
COMMENT     Sequence Label: pRcCMV myc-Plk1 wt (Nigg RG6)
FEATURES             Location/Qualifiers
     source          1..7658
                     /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     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             718..747
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2857..2873)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2857..2873)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2883..2901)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2884..2901)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2921..2938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2927..3151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3197..3710
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3284..3303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3494..3515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3733..3753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3738..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4134..4928
                     /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(4188..4207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4798..4817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5102..5236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5139..5158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5193..5212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5267..5283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5267..5283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5280..5302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5291..5307
                     /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(5315..5321,5322..5339,5340..5345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5360..5381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5498..5515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5669..6257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5749..5768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6428..7219,7220..7288))
                     /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     7051..7070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7289..7393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7468..7487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gcttacattt gcttctgaca caactgtgtt cactagcaac ctcaaacaga caccatggag
      721 cagaagctga tctccgagga ggacctgaac atgaattcct gcagcccggg atccatgagt
      781 gctgcagtga ctgcagggaa gctggcacgg gcaccggccg accctgggaa agccggggtc
      841 cccggagttg cagctcccgg agctccggcg gcggctccac cggcgaaaga gatcccggag
      901 gtcctagtgg acccacgcag ccggcggcgc tatgtgcggg gccgcttttt gggcaagggc
      961 ggctttgcca agtgcttcga gatctcggac gcggacacca aggaggtgtt cgcgggcaag
     1021 attgtgccta agtctctgct gctcaagccg caccagaggg agaagatgtc catggaaata
     1081 tccattcacc gcagcctcgc ccaccagcac gtcgtaggat tccacggctt tttcgaggac
     1141 aacgacttcg tgttcgtggt gttggagctc tgccgccgga ggtctctcct ggagctgcac
     1201 aagaggagga aagccctgac tgagcctgag gcccgatact acctacggca aattgtgctt
     1261 ggctgccagt acctgcaccg aaaccgagtt attcatcgag acctcaagct gggcaacctt
     1321 ttcctgaatg aagatctgga ggtgaaaata ggggattttg gactggcaac caaagtcgaa
     1381 tatgacgggg agaggaagaa gaccctgtgt gggactccta attacatagc tcccgaggtg
     1441 ctgagcaaga aagggcacag tttcgaggtg gatgtgtggt ccattgggtg tatcatgtat
     1501 accttgttag tgggcaaacc accttttgag acttcttgcc taaaagagac ctacctccgg
     1561 atcaagaaga atgaatacag tattcccaag cacatcaacc ccgtggccgc ctccctcatc
     1621 cagaagatgc ttcagacaga tcccactgcc cgcccaacca ttaacgagct gcttaatgac
     1681 gagttcttta cttctggcta tatccctgcc cgtctcccca tcacctgcct gaccattcca
     1741 ccaaggtttt cgattgctcc cagcagcctg gaccccagca accggaagcc cctcacagtc
     1801 ctcaataaag gcttggagaa ccccctgcct gagcgtcccc gggaaaaaga agaaccagtg
     1861 gttcgagaga caggtgaggt ggtcgactgc cacctcagtg acatgctgca gcagctgcac
     1921 agtgtcaatg cctccaagcc ctcggagcgt gggctggtca ggcaagagga ggctgaggat
     1981 cctgcctgca tccccatctt ctgggtcagc aagtgggtgg actattcgga caagtacggc
     2041 cttgggtatc agctctgtga taacagcgtg ggggtgctct tcaatgactc aacacgcctc
     2101 atcctctaca atgatggtga cagcctgcag tacatagagc gtgacggcac tgagtcctac
     2161 ctcaccgtga gttcccatcc caactccttg atgaagaaga tcaccctcct taaatatttc
     2221 cgcaattaca tgagcgagca cttgctgaag gcaggtgcca acatcacgcc gcgcgaaggt
     2281 gatgagctcg cccggctgcc ctacctacgg acctggttcc gcacccgcag cgccatcatc
     2341 ctgcacctca gcaacggcag cgtgcagatc aacttcttcc aggatcacac caagctcatc
     2401 ttgtgcccac tgatggcagc cgtgacctac atcgacgaga agcgggactt ccgcacatac
     2461 cgcctgagtc tcctggagga gtacggctgc tgcaaggagc tggccagccg gctccgctac
     2521 gcccgcacta tggtggacaa gctgctgagc tcacgctcgg ccagcaaccg tctcaaggcc
     2581 tcctaatagc tgccctcccc tccggactgg tgccctcctc actcccacct gcatctgggg
     2641 cccatactgg ttggctcccg cggtgccatg tctgcagtgt gccccccagc cccggtggct
     2701 gggcagagct gcatcatcct tgcaggtggg ggttgctgta taagttattt ttgtacatgt
     2761 tcgggtgtgg gttctacagc cttgtccccc tccccctcaa ccccaccata tgaattgtac
     2821 agaatatttc tattgaattc gagctcggta cccggggatc cactagttct agagggccct
     2881 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     2941 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3001 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3061 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3121 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3181 tcgagggggg atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3241 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3301 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3361 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3421 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3481 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3541 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3601 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaaatattt
     3661 gcttatacaa tcttcctgtt tttggggctt ttctgattat caaccggggt gggtaccgag
     3721 ctcgaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4081 cttggatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4441 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4921 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4981 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5041 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5101 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5161 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5221 ttatcatgtc tggatccgtc gacctcgaga gcttggcgta atcatggtca tagctgtttc
     5281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5401 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5461 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7321 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7381 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7441 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7501 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7561 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     7621 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
