LOCUS       sequence-163997-        7482 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 7482)
  AUTHORS   Jackman M, Kubota Y, den Elzen N, Hagting A, Pines J
  TITLE     Cyclin A- and cyclin E-Cdk complexes shuttle between the nucleus and
            the cytoplasm.
  JOURNAL   Mol Biol Cell. 2002 Mar;13(3):1030-45.
  PUBMED    11907280
REFERENCE   2  (bases 1 to 7482)
  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: "Mol Biol 
            Cell."; date: "2002-03"; volume: "13(3)"; pages: "1030-45"
COMMENT     Sequence Label: pcDNA3/CyclinB1-GFP (28)
FEATURES             Location/Qualifiers
     source          1..7482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2230..2943
                     /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(2257..2285)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2880..2900
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(3034..3052)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3035..3052)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3072..3089)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3078..3302
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3348..3776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3435..3454)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3645..3666
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3785..3805)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3790..4120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3971..4106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4033..4052
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4187..4981
                     /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(4241..4260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4851..4870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5155..5276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5192..5211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5246..5265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5325..5341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5325..5341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5338..5360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5349..5365
                     /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(5373..5379,5380..5397,5398..5403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5418..5439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5556..5573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5727..6315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5807..5826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6486..7277,7278..7346))
                     /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     7109..7128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7347..7451)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatatct
      901 gaggaggacc tttccatggc gctccgagcc accaggaact cgaaaattaa tgctgaaaat
      961 aaggcgaaga tcaacatggc aggcgcaaag cgcgttccta cggcccctgc tgcaacctcc
     1021 aagcccggac tgaggccaag aacagctctt ggggacattg gtaacaaagt cagtgaacaa
     1081 ctgcaggcca aaatgcctat gaagaaggaa gcaaaacctt cagctactgg aaaagtcatt
     1141 gataaaaaac taccaaaacc tcttgaaaag gtacctatgc tggtgccagt gccagtgtct
     1201 gagccagtgc cagagccaga acctgagcca gaacctgagc ctgttaaaga agaaaaactt
     1261 tcgcctgagc ctattttggt tgatactgcc tctccaagcc caatggaaac atctggatgt
     1321 gcccctgcag aagaagacct gtgtcaggct ttctctgatg taattcttgc agtaaatgat
     1381 gtggatgcag aagatggagc tgatccaaac ctttgtagtg aatatgtgaa agatatttat
     1441 gcttatctga gacaacttga ggaagagcaa gcagtcagac caaaatacct actgggtcgg
     1501 gaagtcactg gaaacatgag agccatccta attgactggc tagtacaggt tcaaatgaaa
     1561 ttcaggttgt tgcaggagac catgtacatg actgtctcca ttattgatcg gttcatgcag
     1621 aataattgtg tgcccaagaa gatgctgcag ctggttggtg tcactgccat gtttattgca
     1681 agcaaatatg aagaaatgta ccctccagaa attggtgact ttgcttttgt gactgacaac
     1741 acttatacta agcaccaaat cagacagatg gaaatgaaga ttctaagagc tttaaacttt
     1801 ggtctgggtc ggcctctacc tttgcacttc cttcggagag catctaagat tggagaggtt
     1861 gatgtcgagc aacatacttt ggccaaatac ctgatggaac taactatgtt ggactatgac
     1921 atggtgcact ttcctccttc tcaaattgca gcaggagctt tttgcttagc actgaaaatt
     1981 ctggataatg gtgaatggac accaactcta caacattacc tgtcatatac tgaagaatct
     2041 cttcttccag ttatgcagca cctggctaag aatgtagtca tggtaaatca aggacttaca
     2101 aagcacatga ctgtcaagaa caagtatgcc acatcgaagc atgctaagat cagcactcta
     2161 ccacagctga attctgcact agttcaagat ttagccaagg ctgtggcaaa ggtggcagga
     2221 gcgcaattca gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
     2281 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
     2341 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     2401 acacttgtca ctactctcac ttatggtgtt caatgctttt caagatatcc agatcatatg
     2461 aagcggcacg acttcttcaa gagcgccatg cctgagggat acgtgcagga gaggaccatc
     2521 ttcttcaagg acgacgggaa ctacaagaca cgtgctgaag tcaagtttga gggagacacc
     2581 ctcgtcaaca ggatcgagct taagggaatc gatttcaagg aggacggaaa catcctcggc
     2641 cacaagttgg aatacaacta caactcccac aacgtataca tcatggccga caagcaaaag
     2701 aacggcatca aagccaactt caagacccgc cacaacatcg aagacggcgg cgtgcaactc
     2761 gctgatcatt atcaacaaaa tactccaatt ggcgatgacc ctgtcctttt accagacaac
     2821 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     2881 gtccttcttg agtttgtaac ggctgctggg attacacatg gcatggatga actatacaaa
     2941 taggatccac tagtaacggc cgccagtgtg ctggaattct gcagatatcc atcacactgg
     3001 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3061 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3121 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3181 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3241 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3301 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3361 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3421 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3481 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3541 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3601 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3661 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3721 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3781 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     3841 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3961 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4021 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4081 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4141 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5281 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7441 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
