LOCUS       sequence-409983-        6009 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 6009)
  AUTHORS   Alberts AS
  TITLE     Identification of a carboxyl-terminal diaphanous-related formin 
            homology protein autoregulatory domain.
  JOURNAL   J Biol Chem. 2001 Jan 26. 276(4):2824-30.
  PUBMED    11035012
REFERENCE   2  (bases 1 to 6009)
  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 Biol 
            Chem. 2001 Jan 26. 276(4):2824-30."
COMMENT     Sequence Label: pEGFPm-DAD
FEATURES             Location/Qualifiers
     source          1..6009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2336..3514
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2567..3505
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3462..3482
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3586..3603
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3613..3642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3658..4365
                     /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
                     FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     primer_bind     complement(3685..3713)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4308..4328
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4910..5304
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(5615..5632)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5615..5631)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5624..5646)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5840..5859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5959..5981
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 acatgattac gaattaattc gagctcgccc agctttgcaa agatggataa agttttaaac
     2281 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
     2341 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2401 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2461 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2521 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     2581 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     2641 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     2701 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     2761 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     2821 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     2881 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     2941 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     3001 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     3061 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     3121 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     3181 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     3241 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     3301 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     3361 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     3421 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     3481 tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt agcttgctta catttgcttc
     3541 tgacacaact gtgttcacta gcaacctcaa acagacacca tgccgcatca ccaccatcat
     3601 cacgccatgc cggagcagaa gctgatatcc gaggaggacc tggccatggc cggatctagt
     3661 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     3721 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     3781 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     3841 actctcactt atggtgttca atgcttttca agatacccag atcatatgaa gcggcacgac
     3901 ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga ggaccatctt cttcaaggac
     3961 gacgggaact acaagacacg tgctgaagtc aagtttgagg gagacaccct cgtcaacagg
     4021 atcgagctta agggaatcga tttcaaggag gacggaaaca tcctcggcca caagttggaa
     4081 tacaactaca actcccacaa cgtatacatc atggccgaca agcaaaagaa cggcatcaaa
     4141 gccaacttca agacccgcca caacatcgaa gacggcggcg tgcaactcgc tgatcattat
     4201 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     4261 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     4321 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacggatc catgaaaact
     4381 gagggagatg agacaggagt gatggatagt ctgctggagg ccttgcagtc aggggctgcc
     4441 ttccgcgaca gaagaaaaag gacaccaaag ctgaaagata ttcggcagag tctcagcccg
     4501 atgtctcaga ggcctgttct caaagtttgt aaccatgaaa atcagaaaat gccgttgaca
     4561 gaagggtcac gtccacacca cagtatcaat tgcaactcca ccaggactcc agtcgccaag
     4621 gagcttaatt ataatctaga cactcatgcg tctacaggga ggatcaaggc agttgagaag
     4681 gaagcctgta atgcagaaag caacaaaaaa aaggaaatgg aacttcttgg ctctgttgct
     4741 aaaagcgaat cagttcctga agttgaagcc ctgctggcaa gattacgagc tttataagaa
     4801 ttcctcgaga tcgattagac tagtctagaa attcacccca ccagtgcagg ctgcctatca
     4861 gaaagtggtg gctggtgtgg ctaatgccct ggcccacaag tatcactaag ctcgctttct
     4921 tgctgtccaa tttctattaa aggttccttt gttccctaag tccaactact aaactggggg
     4981 atattatgaa gggccttgag catctggatt ctgcctaata aaaaacattt attttcattg
     5041 caatgatgta tttaaattat ttctgaatat tttactaaaa agggaatgtg ggaggtcagt
     5101 gcatttaaaa cataaagaaa tgaagagcta gttcaaacct tgggaaaata cactatatct
     5161 taaactccat gaaagaaggt gaggctgcaa acagctaatg cacattggca acagcccctg
     5221 atgcctatgc cttattcatc cctcagaaaa ggattcaagt agaggcttga tttggaggtt
     5281 aaagttttgc tatgctgtat tttacattac ttattgtttt agctgtcctc atgaatgtct
     5341 tttcactacc catttgctta tcctgcatct ctcagccttg actccactca gttctcttgc
     5401 ttagagatac cacctttccc ctgaagtgtt ccttccatgt tttacggcga gatggtttct
     5461 cctcgcctgg ccactcagcc ttagttgtct ctgttgtctt atagaggtct acttgaagaa
     5521 ggaaaaacag ggggcatggt ttgactgtcc tgtgagccct tcttccctgc ctcccccact
     5581 cacagtgacc cggaatctgc agcccaagct tggcactggc cgtcgtttta caacgtcgtg
     5641 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     5701 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5761 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5821 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5881 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5941 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6001 aacgcgcga
//
