LOCUS       sequence-152165-        7477 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For mammalian expression of human MKL1 with a 3x FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  AUTHORS   Cen B, Selvaraj A, Burgess RC, Hitzler JK, Ma Z, Morris SW, Prywes R
  TITLE     Megakaryoblastic leukemia 1, a potent transcriptional coactivator 
            for serum response factor (SRF), is required for serum induction of 
            SRF target genes.
  JOURNAL   Mol Cell Biol. 2003 Sep . 23(18):6597-608.
  PUBMED    12944485
REFERENCE   2  (bases 1 to 7477)
  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 Cell 
            Biol. 2003 Sep . 23(18):6597-608."
COMMENT     Sequence Label: p3xFLAG-MKL1
FEATURES             Location/Qualifiers
     source          1..7477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..97
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     89..106
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     90..106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        267..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        647..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             880..945
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1300..1311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3767..4389
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4413..4433)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4418..4747
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4598..4733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4785..4804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4786..4804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4818..4834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4818..4834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4831..4853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4842..4858
                     /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(4866..4872,4873..4890,4891..4896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4911..4932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5049..5066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5300..5319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6971..7426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7058..7077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7268..7289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
       61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgatcta
      121 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
      181 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
      241 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      661 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct cgtttagtga accgtcagaa ttaaccatgg actacaaaga ccatgacggt
      901 gattataaag atcatgacat cgattacaag gatgacgatg acaagcttgc ggccgcgaat
      961 tcgatgccgc ctttgaaaag tccagccgca tttcatgagc agagaaggag cttggagcgg
     1021 gccaggacag aggactatct caaacggaag attcgttccc ggccggagag atcggagctg
     1081 gtcaggatgc acattttgga agagacctcg gctgagccat ccctccaggc caagcagctg
     1141 aagctgaaga gagccagact agccgatgac ctcaatgaga agattgcaca gaggcctggc
     1201 cccatggagc tggtggagaa gaacatcctt cctgttgagt ccagcctgaa ggaagccatc
     1261 attgtgggcc aggtgaacta tcccaaagta gcagacagct cttccttcga tgaggacagc
     1321 agcgatgcct tatcccccga gcagcctgcc agccatgagt cccagggttc tgtgccgtca
     1381 cccctggagg cccgagtcag cgaaccactg ctcagtgcca cctctgcatc ccccacccag
     1441 gttgtgtctc aacttccgat gggccgggat tccagagaaa tgcttttcct ggcagagcag
     1501 cctcctctgc ctcccccacc tctgctgcct cccagcctca ccaatggaac cactatcccc
     1561 actgccaagt ccacccccac actcattaag caaagccaac ccaagtctgc cagtgagaag
     1621 tcacagcgca gcaagaaggc caaggagctg aagccaaagg tgaagaagct caagtaccac
     1681 cagtacatcc ccccggacca gaagcaggac aggggggcac cccccatgga ctcatcctac
     1741 gccaagatcc tgcagcagca gcagctcttc ctccagctgc agatcctcaa ccagcagcag
     1801 cagcagcacc acaactacca ggccatcctg cctgccccgc caaagtcagc aggcgaggcc
     1861 ctgggaagca gcgggacccc cccagtacgc agcctctcca ctaccaatag cagctccagc
     1921 tcgggcgccc ctgggccctg tgggctggca cgtcagaaca gcacctcact gactggcaag
     1981 ccgggagccc tgccggccaa cctggacgac atgaaggtgg cagagctgaa gcaggagctg
     2041 aagttgcgat cactgcctgt ctcgggcacc aaaactgagc tgattgagcg ccttcgagcc
     2101 tatcaagacc aaatcagccc tgtgccagga gcccccaagg cccctgccgc cacctctatc
     2161 ctgcacaagg ctggcgaggt ggtggtagcc ttcccagcgg cccggctgag cacggggcca
     2221 gccctggtgg cagcaggcct ggctccagct gaggtggtgg tggccacggt ggccagcagt
     2281 ggggtggtga agtttggcag cacgggctcc acgccccccg tgtctcccac cccctcggag
     2341 cgctcactgc tcagcacggg cgatgaaaac tccacccccg gggacacctt tggtgagatg
     2401 gtgacatcac ctctgacgca gctgaccctg caggcctcgc cactgcagat cctcgtgaag
     2461 gaggagggcc cccgggccgg gtcctgttgc ctgagccctg gggggcgggc ggagctagag
     2521 gggcgcgaca aggaccagat gctgcaggag aaagacaagc agatcgaggc gctgacgcgc
     2581 atgctccggc agaagcagca gctggtggag cggctcaagc tgcagctgga gcaggagaag
     2641 cgagcccagc agcccgcccc cgcccccgcc cccctcggca cccccgtgaa gcaggagagc
     2701 agcttctcca gctgccagct gagccagcag cccctgggcc ccgctcaccc attcaacccc
     2761 agcctggcgg ccccagccac caaccacata gacccttgtg ctgtggcccc ggggcccccg
     2821 tccgtggtgg tgaagcagga agccttgcag cctgagcccg agccggtccc cgccccccag
     2881 ttgcttctgg ggcctcaggg ccccagcctc atcaaggggg ttgcacctcc caccctcatc
     2941 accgactcca cagggaccca ccttgtcctc accgtgacca ataagaatgc agacagccct
     3001 ggcctgtcca gtgggagccc ccagcagccc tcgtcccagc ctggctctcc agcgcctgcc
     3061 ccctctgccc agatggacct ggagcaccca ctgcagcccc tctttggggc ccccacttct
     3121 ctgctgaaga aggaaccacc tggctatgag gaagccatga gccagcagcc caaacagcag
     3181 gaaaatggtt cctcaagcca gcagatggac gacctgtttg acattctcat tcagagcgga
     3241 gaaatttcag cagatttcaa agagccgcca tccctgccag ggaaggagaa gccatccccg
     3301 aagacagtct gtgggtcccc cctggcagca cagccatcac cttctgctga gctcccccag
     3361 gctgccccac ctcctccagg ctcaccctcc ctccctggac gcctggagga cttcctggag
     3421 agcagcacgg ggctgcccct gctgaccagt gggcatgacg ggccagagcc cctttccctc
     3481 attgacgacc tccatagcca gatgctgagc agcactgcca tcctggacca ccccccgtca
     3541 cccatggaca cctcggaatt gcactttgtt cctgagccca gcagcaccat gggcctggac
     3601 ctggctgatg gccacctgga cagcatggac tggctggagc tgtcgtcagg tggtcccgtg
     3661 ctgagcctag cccccctcag caccacagcc cccagcctct tctccacaga cttcctcgat
     3721 ggccatgatt tgcagctgca ctgggattcc tgcttgtagg gatcccgggt ggcatccctg
     3781 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3841 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3901 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3961 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     4021 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     4081 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4141 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4201 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     4261 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     4321 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     4381 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4501 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4561 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4621 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4681 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4741 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     4801 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4861 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4921 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7021 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7081 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7141 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7201 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7261 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7321 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7381 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     7441 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggc
//
