LOCUS       sequence-163995-        7470 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 7470)
  AUTHORS   den Elzen N, Pines J
  TITLE     Cyclin A is destroyed in prometaphase and can delay chromosome 
            alignment and anaphase.
  JOURNAL   J Cell Biol. 2001 Apr 2;153(1):121-36.
  PUBMED    11285279
REFERENCE   2  (bases 1 to 7470)
  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 
            Biol."; date: "2001-04-2"; volume: "153(1)"; pages: "121-36"
COMMENT     Sequence Label: pcDNA3-myc-Delta-D-box-CycA-GFP (525)
FEATURES             Location/Qualifiers
     source          1..7470
                     /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             901..930
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2218..2931
                     /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="SKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2245..2273)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2868..2888
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(3022..3040)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3023..3040)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3060..3077)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3066..3290
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3336..3764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3423..3442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3633..3654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3773..3793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3778..4108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3959..4094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4021..4040
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4175..4969
                     /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(4229..4248)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4839..4858
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5143..5264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5180..5199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5234..5253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5313..5329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5313..5329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5326..5348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5337..5353
                     /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(5361..5367,5368..5385,5386..5391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5406..5427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5544..5561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5715..6303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5795..5814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6474..7265,7266..7334))
                     /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     7097..7116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7335..7439)
                     /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 gcttatcatg
      901 gagcaaaaac ttatatctga ggaggacctt tccatggtgg gcaactctgc gccggggcct
      961 gcgacccgcg aggcgggctc ggcgctgcta gcattgcagc agacggcgct ccaagaggac
     1021 caggagaata tcaacccgga aaaggcagcg cccgtccaac aaccacggac cgggaacccg
     1081 cggggtctag cgcagcagca gaggccgaag acgagacggg ttgcacccct taaggatctt
     1141 cctgtaaatg atgagcatgt caccgttcct ccttggaaag caaacagtaa acagcctgcg
     1201 ttcaccattc atgtggatga agcagaaaaa gaagctcaga agaagccagc tgaatctcaa
     1261 aaaatagagc gtgaagatgc cctggctttt aattcagcca ttagtttacc tggacccaga
     1321 aaaccattgg tccctcttga ttatccaatg gatggtagtt ttgagtcacc acatactatg
     1381 gacatgtcaa ttgtattaga agatgaaaag ccagtgagtg ttaatgaagt accagactac
     1441 catgaggata ttcacacata ccttagggaa atggaggtta aatgtaaacc taaagtgggt
     1501 tacatgaaga aacagccaga catcactaac agtatgagag ctatcctcgt ggactggtta
     1561 gttgaagtag gagaagaata taaactacag aatgagaccc tgcatttggc tgtgaactac
     1621 attgataggt tcctgtcttc catgtcagtg ctgagaggaa aacttcagct tgtgggcact
     1681 gctgctatgc tgttagcctc aaagtttgaa gaaatatacc ccccagaagt agcagagttt
     1741 gtgtacatta cagatgatac ctacaccaag aaacaagttc tgagaatgga gcatctagtt
     1801 ttgaaagtcc ttacttttga cttagctgct ccaacagtaa atcagtttct tacccaatac
     1861 tttctgcatc agcagcctgc aaactgcaaa gttgaaagtt tagcaatgtt tttgggagaa
     1921 ttaagtttga tagatgctga cccatacctc aagtatttgc catcagttat tgctggagct
     1981 gcctttcatt tagcactcta cacagtcacg ggacaaagct ggcctgaatc attaatacga
     2041 aagactggat ataccctgga aagtcttaag ccttgtctca tggaccttca ccagacctac
     2101 ctcaaagcac cacagcatgc acaacagtca ataagagaaa agtacaaaaa ttcaaagtat
     2161 catggtgttt ctctcctcaa cccaccagag acactaaatc tggcaggagc ggaattcagt
     2221 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     2281 aatgggtaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     2341 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     2401 actctcactt atggtgttca atgcttttca agatatccag atcatatgaa gcggcacgac
     2461 ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga ggaccatctt cttcaaggac
     2521 gacgggaact acaagacacg tgctgaagtc aagtttgagg gagacaccct cgtcaacagg
     2581 atcgagctta agggaatcga tttcaaggag gacggaaaca tcctcggcca caagttggaa
     2641 tacaactaca actcccacaa cgtatacatc atggccgaca agcaaaagaa cggcatcaaa
     2701 gccaacttca agacccgcca caacatcgaa gacggcggcg tgcaactcgc tgatcattat
     2761 caacaaaata ctccaattgg cgatgaccct gtccttttac cagacaacca ttacctgtcc
     2821 acacaatccg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2881 tttgtaacgg ctgctgggat tacacatggc atggatgaac tatacaaata ggatccacta
     2941 gtaacggccg ccagtgtgct ggaattctgc agatatccat cacactggcg gccgctcgag
     3001 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
     3061 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3121 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3181 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3241 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3301 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3361 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3421 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3481 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3541 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3601 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3661 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3721 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3781 tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3901 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3961 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4021 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4081 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4141 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4201 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4261 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4321 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4381 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4441 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4501 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4561 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4621 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4681 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4741 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4801 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4861 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4921 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4981 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5041 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5101 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5161 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5221 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5281 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5341 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5401 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5461 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5521 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5881 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6121 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6361 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7441 acatttcccc gaaaagtgcc acctgacgtc
//
