LOCUS       sequence-421230-        6425 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 6425)
  AUTHORS   Biggs JR, Peterson LF, Zhang Y, Kraft AS, Zhang DE
  TITLE     AML1/RUNX1 phosphorylation by cyclin-dependent kinases regulates the
            degradation of AML1/RUNX1 by the anaphase-promoting complex.
  JOURNAL   Mol Cell Biol. 2006 Oct . 26(20):7420-9.
  PUBMED    17015473
REFERENCE   2  (bases 1 to 6425)
  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: "Mol Cell 
            Biol. 2006 Oct . 26(20):7420-9."
COMMENT     Sequence Label: pCMV-3xFlag-ETO
FEATURES             Location/Qualifiers
     source          1..6425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(48..66)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(80..96)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(80..96)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(93..115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    104..120
                     /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(128..134,135..152,153..158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    173..194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(311..328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(482..1070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(562..581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1241..2032,2033..2101))
                     /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     1864..1883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2102..2206)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2233..2688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2320..2339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2530..2551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2814..2836
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2828..2845
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2829..2845
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3006..3385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3386..3589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3540..3560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3619..3684
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5454..6076
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5653..5672)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6100..6120)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6105..6425,1..9)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6285..6420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc
       61 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      121 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      181 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      241 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2221 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2521 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2581 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2641 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2701 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2761 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2821 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2881 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2941 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     3001 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     3061 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3241 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3301 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3601 ccgtcagaat taaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc
     3661 gattacaagg atgacgatga caagcttgcg gcccattcaa ggatgcctga tcgtactgag
     3721 aagcactcca caatgccaga ctcacctgtg gatgtgaaga cgcaatctag gctgactcct
     3781 ccaacaatgc cacctccccc aactactcaa ggagctccaa gaaccagttc atttacaccg
     3841 acaacgttaa ctaatggcac gagccattct cctacagcct tgaatggcgc cccctcacca
     3901 cccaatggct tcagcaatgg gccttcctct tcttcctcct cctctctggc taatcaacag
     3961 ctgcccccag cctgtggtgc caggcaactc agcaagctga aaaggttcct tactaccctg
     4021 cagcagtttg gcaatgacat ttcacccgag ataggagaaa gagttcgcac cctcgttctg
     4081 ggactagtga actccacttt gacaattgaa gaatttcatt ccaaactgca agaagctact
     4141 aacttcccac tgagaccttt tgtcatccca tttttgaagg ccaacttgcc cctgctgcag
     4201 cgtgagctcc tccactgcgc aagactggcc aaacagaacc ctgcccagta cctcgcccag
     4261 catgaacagc tgcttctgga tgccagcacc acctcacctg ttgactcctc agagctgctt
     4321 ctcgatgtga acgaaaacgg gaagaggcga actccagaca gaaccaaaga aaatggcttt
     4381 gacagagagc ctttgcactc agaacatcca agcaagcgac catgcactat tagcccaggc
     4441 cagcggtaca gtccaaataa cggcttatcc taccagccca atggcctgcc tcaccctacc
     4501 ccacctccac ctcagcatta ccgtttggat gatatggcca ttgcccacca ctacagggac
     4561 tcctatcgac accccagcca cagggacctc agggacagaa acagacctat ggggttgcat
     4621 ggcacacgtc aagaagaaat gattgatcac agactaacag acagagaatg ggcagaagag
     4681 tggaaacatc ttgaccatct gttaaactgc ataatggaca tggtagaaaa aacaaggcga
     4741 tctctcaccg tactaaggcg gtgtcaagaa gcagaccggg aagaattgaa ttactggatc
     4801 cggcggtaca gtgacgccga ggacttaaaa aaaggtggcg gcagtagcag cagccactct
     4861 aggcagcaga gtcccgtcaa cccagaccca gttgcactag acgcgcatcg ggaattcctt
     4921 cacaggcctg cgtctggata cgtgccagag gagatctgga agaaagctga ggaggccgtc
     4981 aatgaggtga agcgccaggc gatgacggag ctgcagaagg ccgtgtctga ggcggagcgg
     5041 aaagcccacg acatgatcac aacagagagg gccaagatgg agcgcacggt cgccgaggcc
     5101 aaacggcagg cggcggagga cgcactggca gttatcaatc agcaggagga ttcaagcgag
     5161 agttgctgga attgtggccg taaagcgagt gaaacctgca gtggctgtaa cacagcccga
     5221 tactgtggct cattttgcca gcacaaagac tgggagaagc accatcacat ctgtggacag
     5281 accctgcagg cccagcagca gggagacaca cctgcagtca gctcctctgt cacgcccaac
     5341 agcggggctg ggagcccgat ggacacacca ccagcagcca ctccgaggtc aaccaccccg
     5401 ggaacccctt ccaccataga gacaacccct cgctagtgac tctagaggat cccgggtggc
     5461 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     5521 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     5581 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     5641 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     5701 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     5761 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     5821 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     5881 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa
     5941 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac
     6001 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
     6061 agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt
     6121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6181 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6241 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6301 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6361 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6421 taggc
//
