LOCUS       sequence-270150-        8201 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 8201)
  AUTHORS   Muehlich S, Wang R, Lee SM, Lewis TC, Dai C, Prywes R
  TITLE     Serum-induced phosphorylation of the serum response factor 
            coactivator MKL1 by the extracellular signal-regulated kinase 1/2 
            pathway inhibits its nuclear localization.
  JOURNAL   Mol Cell Biol. 2008 Oct;28(20):6302-13. doi: 10.1128/MCB.00427-08. 
            Epub 2008 Aug 11.
  PUBMED    18694962
REFERENCE   2  (bases 1 to 8201)
  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"; doi: 
            "10.1128/MCB.00427-08"; journalName: "Mol Cell Biol"; date: 
            "2008-10"; volume: "28"; issue: "20"; pages: "6302-13"
COMMENT     Sequence Label: pCin4-3xFLAG-MKL1
FEATURES             Location/Qualifiers
     source          1..8201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        28..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     656..675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        656..674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        708..759
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     CDS             789..854
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     intron          3736..3965
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3948..3967
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4148..4610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4204..4221)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4431..4450
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4646..5449
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4709..4728)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5319..5338
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5481..5498)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5487..5711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5758..5777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5837..5853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5837..5853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5850..5872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5861..5877
                     /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(5885..5891,5892..5909,5910..5915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5930..5951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6068..6085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6239..6827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6319..6338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6998..7789,7790..7858))
                     /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     7621..7640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7859..7963)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8038..8057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      661 cgactcacta tagggagacc caagcttggt accgagctcg gatcgatgac gcaaatgggc
      721 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagaat
      781 taaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg
      841 atgacgatga caagcttgcg gccgcgaatt cgatgccgcc tttgaaaagt ccagccgcat
      901 ttcatgagca gagaaggagc ttggagcggg ccaggacaga ggactatctc aaacggaaga
      961 ttcgttcccg gccggagaga tcggagctgg tcaggatgca cattttggaa gagacctcgg
     1021 ctgagccatc cctccaggcc aagcagctga agctgaagag agccagacta gccgatgacc
     1081 tcaatgagaa gattgcacag aggcctggcc ccatggagct ggtggagaag aacatccttc
     1141 ctgttgagtc cagcctgaag gaagccatca ttgtgggcca ggtgaactat cccaaggtcg
     1201 cggattcaag tagtttcgat gaggacagca gcgatgcctt atcccccgag cagcctgcca
     1261 gccatgagtc ccagggttct gtgccgtcac ccctggaggc ccgagtcagc gaaccactgc
     1321 tcagtgccac ctctgcatcc cccacccagg ttgtgtctca acttccgatg ggccgggatt
     1381 ccagagaaat gcttttcctg gcagagcagc ctcctctgcc tcccccacct ctgctgcctc
     1441 ccagcctcac caatggaacc actatcccca ctgccaagtc cacccccaca ctcattaagc
     1501 aaagccaacc caagtctgcc agtgagaagt cacagcgcag caagaaggcc aaggagctga
     1561 agccaaaggt gaagaagctc aagtaccacc agtacatccc cccggaccag aagcaggaca
     1621 ggggggcacc ccccatggac tcatcctacg ccaagatcct gcagcagcag cagctcttcc
     1681 tccagctgca gatcctcaac cagcagcagc agcagcacca caactaccag gccatcctgc
     1741 ctgccccgcc aaagtcagca ggcgaggccc tgggaagcag cgggaccccc ccagtacgca
     1801 gcctctccac taccaatagc agctccagct cgggcgcccc tgggccctgt gggctggcac
     1861 gtcagaacag cacctcactg actggcaagc cgggagccct gccggccaac ctggacgaca
     1921 tgaaggtggc agagctgaag caggagctga agttgcgatc actgcctgtc tcgggcacca
     1981 aaactgagct gattgagcgc cttcgagcct atcaagacca aatcagccct gtgccaggag
     2041 cccccaaggc ccctgccgcc acctctatcc tgcacaaggc tggcgaggtg gtggtagcct
     2101 tcccagcggc ccggctgagc acggggccag ccctggtggc agcaggcctg gctccagctg
     2161 aggtggtggt ggccacggtg gccagcagtg gggtggtgaa gtttggcagc acgggctcca
     2221 cgccccccgt gtctcccacc ccctcggagc gctcactgct cagcacgggc gatgaaaact
     2281 ccacccccgg ggacaccttt ggtgagatgg tgacatcacc tctgacgcag ctgaccctgc
     2341 aggcctcgcc actgcagatc ctcgtgaagg aggagggccc ccgggccggg tcctgttgcc
     2401 tgagccctgg ggggcgggcg gagctagagg ggcgcgacaa ggaccagatg ctgcaggaga
     2461 aagacaagca gatcgaggcg ctgacgcgca tgctccggca gaagcagcag ctggtggagc
     2521 ggctcaagct gcagctggag caggagaagc gagcccagca gcccgccccc gcccccgccc
     2581 ccctcggcac ccccgtgaag caggagagca gcttctccag ctgccagctg agccagcagc
     2641 ccctgggccc cgctcaccca ttcaacccca gcctggcggc cccagccacc aaccacatag
     2701 acccttgtgc tgtggccccg gggcccccgt ccgtggtggt gaagcaggaa gccttgcagc
     2761 ctgagcccga gccggtcccc gccccccagt tgcttctggg gcctcagggc cccagcctca
     2821 tcaagggggt tgcacctccc accctcatca ccgactccac agggacccac cttgtcctca
     2881 ccgtgaccaa taagaatgca gacagccctg gcctgtccag tgggagcccc cagcagccct
     2941 cgtcccagcc tggctctcca gcgcctgccc cctctgccca gatggacctg gagcacccac
     3001 tgcagcccct ctttggggcc cccacttctc tgctgaagaa ggaaccacct ggctatgagg
     3061 aagccatgag ccagcagccc aaacagcagg aaaatggttc ctcaagccag cagatggacg
     3121 acctgtttga cattctcatt cagagcggag aaatttcagc agatttcaaa gagccgccat
     3181 ccctgccagg gaaggagaag ccatccccga agacagtctg tgggtccccc ctggcagcac
     3241 agccatcacc ttctgctgag ctcccccagg ctgccccacc tcctccaggc tcaccctccc
     3301 tccctggacg cctggaggac ttcctggaga gcagcacggg gctgcccctg ctgaccagtg
     3361 ggcatgacgg gccagagccc ctttccctca ttgacgacct ccatagccag atgctgagca
     3421 gcactgccat cctggaccac cccccgtcac ccatggacac ctcggaattg cactttgttc
     3481 ctgagcccag cagcaccatg ggcctggacc tggctgatgg ccacctggac agcatggact
     3541 ggctggagct gtcgtcaggt ggtcccgtgc tgagcctagc ccccctcagc accacagccc
     3601 ccagcctctt ctccacagac ttcctcgatg gccatgattt gcagctgcac tgggattcct
     3661 gcttgtaggg atccactagt aacggccgcc agtgtgctgg aattaattcg ctgtctgcga
     3721 gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat
     3781 tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga
     3841 gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg
     3901 gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc
     3961 cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg cggccaattc
     4021 cgcccctctc ccccccccnn nnnnnnnccc ccccccctaa cgttactggc cgaagccgct
     4081 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4141 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4201 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     4261 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     4321 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     4381 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     4441 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     4501 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     4561 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca
     4621 acccgggata attcctgcag ccaatatggg atcggccatt gaacaagatg gattgcacgc
     4681 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4741 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4801 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4861 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4921 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4981 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5041 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5101 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5161 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg
     5221 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5281 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5341 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5401 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gggatcaatt
     5461 ctctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5521 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5581 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5641 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5701 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgagtgcat tctagttgtg
     5761 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5821 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6001 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6061 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6961 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7021 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7081 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7141 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7201 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7261 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7321 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7381 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7441 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7501 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7561 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgtccggcg tcaatacggg
     7621 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7681 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7741 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7801 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7861 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7921 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7981 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     8041 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     8101 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     8161 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt c
//
