LOCUS       sequence-246247-        6399 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  AUTHORS   Hagting A, Jackman M, Simpson K, Pines J
  TITLE     Translocation of cyclin B1 to the nucleus at prophase requires a 
            phosphorylation-dependent nuclear import signal.
  JOURNAL   Curr Biol. 1999 Jul 1. 9(13):680-9.
  PUBMED    10395539
REFERENCE   2  (bases 1 to 6399)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Biol. 
            1999 Jul 1. 9(13):680-9."
COMMENT     Sequence Label: pCMX/cyclinB1-GFP (27)
FEATURES             Location/Qualifiers
     source          1..6399
                     /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             1996..2709
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2023..2051)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2646..2666
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          2816..2881
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3011..3031
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3303..3437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3340..3359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3394..3413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3895..4030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4095..4112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5025..5816,5817..5885))
                     /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     5648..5667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5886..5990)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6058..6076
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6114..6136)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6236..6255
                     /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 atatctgagg aggacctttc catggcgctc cgagccacca ggaactcgaa aattaatgct
      721 gaaaataagg cgaagatcaa catggcaggc gcaaagcgcg ttcctacggc ccctgctgca
      781 acctccaagc ccggactgag gccaagaaca gctcttgggg acattggtaa caaagtcagt
      841 gaacaactgc aggccaaaat gcctatgaag aaggaagcaa aaccttcagc tactggaaaa
      901 gtcattgata aaaaactacc aaaacctctt gaaaaggtac ctatgctggt gccagtgcca
      961 gtgtctgagc cagtgccaga gccagaacct gagccagaac ctgagcctgt taaagaagaa
     1021 aaactttcgc ctgagcctat tttggttgat actgcctctc caagcccaat ggaaacatct
     1081 ggatgtgccc ctgcagaaga agacctgtgt caggctttct ctgatgtaat tcttgcagta
     1141 aatgatgtgg atgcagaaga tggagctgat ccaaaccttt gtagtgaata tgtgaaagat
     1201 atttatgctt atctgagaca acttgaggaa gagcaagcag tcagaccaaa atacctactg
     1261 ggtcgggaag tcactggaaa catgagagcc atcctaattg actggctagt acaggttcaa
     1321 atgaaattca ggttgttgca ggagaccatg tacatgactg tctccattat tgatcggttc
     1381 atgcagaata attgtgtgcc caagaagatg ctgcagctgg ttggtgtcac tgccatgttt
     1441 attgcaagca aatatgaaga aatgtaccct ccagaaattg gtgactttgc ttttgtgact
     1501 gacaacactt atactaagca ccaaatcaga cagatggaaa tgaagattct aagagcttta
     1561 aactttggtc tgggtcggcc tctacctttg cacttccttc ggagagcatc taagattgga
     1621 gaggttgatg tcgagcaaca tactttggcc aaatacctga tggaactaac tatgttggac
     1681 tatgacatgg tgcactttcc tccttctcaa attgcagcag gagctttttg cttagcactg
     1741 aaaattctgg ataatggtga atggacacca actctacaac attacctgtc atatactgaa
     1801 gaatctcttc ttccagttat gcagcacctg gctaagaatg tagtcatggt aaatcaagga
     1861 cttacaaagc acatgactgt caagaacaag tatgccacat cgaagcatgc taagatcagc
     1921 actctaccac agctgaattc tgcactagtt caagatttag ccaaggctgt ggcaaaggtg
     1981 gcaggagcgc aattcagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     2041 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     2101 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     2161 tggccaacac ttgtcactac tctcacttat ggtgttcaat gcttttcaag atatccagat
     2221 catatgaagc ggcacgactt cttcaagagc gccatgcctg agggatacgt gcaggagagg
     2281 accatcttct tcaaggacga cgggaactac aagacacgtg ctgaagtcaa gtttgaggga
     2341 gacaccctcg tcaacaggat cgagcttaag ggaatcgatt tcaaggagga cggaaacatc
     2401 ctcggccaca agttggaata caactacaac tcccacaacg tatacatcat ggccgacaag
     2461 caaaagaacg gcatcaaagc caacttcaag acccgccaca acatcgaaga cggcggcgtg
     2521 caactcgctg atcattatca acaaaatact ccaattggcg atgaccctgt ccttttacca
     2581 gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac
     2641 cacatggtcc ttcttgagtt tgtaacggct gctgggatta cacatggcat ggatgaacta
     2701 tacaaatagg atcctggcca gctagctagg tagctagagg atctttgtga aggaacctta
     2761 cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa
     2821 tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
     2881 gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa
     2941 cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca
     3001 ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt
     3061 gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta
     3121 caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat atttgatgta
     3181 tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3241 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3301 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3361 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3421 cttatcatgt ctggatcagc ttcagaagat ggcggagggc ctccaacaca gtaattttcc
     3481 tcccgactct taaaatagaa aatgtcaagt cagttaagca ggaagtgact aactgaccgc
     3541 agctggccgt gcgacatcct cttttaatta gttgctaggc aactgccctc cagagggcag
     3601 tgtggttttg caagaggaag caaaaagcct ctccacccag gcctagaatg tttccaccca
     3661 atcattacta tgacaacagc tgtttttttt agtattaagc agaggccggg ggcccctggc
     3721 ctccgcttac tctggagaaa aagaagagag gcattgtaga ggcttccaga ggcaacttgt
     3781 caaaacagga ctgcttctat ttctgtcaca ctgtctggcc ctgtcacaag gtccagcacc
     3841 tccatacccc ctttaataag cagtttggga acgggtgcgg gtcttactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggsc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4081 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4981 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5101 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5161 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5221 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5281 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5341 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5401 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5461 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5521 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5581 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5641 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5701 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5761 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5821 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5881 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6001 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6061 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6121 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6181 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6241 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     6301 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6361 cgatcggtgc gggcctcttc gctattacgc cagcgcgtt
//
