LOCUS       sequence-490275-        7320 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Expressing constitutively active mimetic of phospho-Thr508 LIM 
            Kinase 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7320)
  AUTHORS   Edwards DC, Gill GN
  TITLE     Structural features of LIM kinase that control effects on the actin 
            cytoskeleton.
  JOURNAL   J Biol Chem. 1999 Apr 16;274(16):11352-61. doi: 
            10.1074/jbc.274.16.11352.
  PUBMED    10196227
REFERENCE   2  (bases 1 to 7320)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.274.16"; journalName: "J Biol Chem"; date: "1999-04-16-
            16"; volume: "274"; issue: "16"; pages: "11352-61"
COMMENT     Sequence Label: LIMK1 T508EE
FEATURES             Location/Qualifiers
     source          1..7320
                     /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"
     primer_bind     complement(2911..2928)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2917..3141
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3187..3615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3274..3293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3484..3505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3624..3644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3629..3958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3809..3944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3871..3890
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4025..4819
                     /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(4079..4098)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4689..4708
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4993..5114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5030..5049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5084..5103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5163..5179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5163..5179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5176..5198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5187..5203
                     /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(5211..5217,5218..5235,5236..5241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5256..5277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5394..5411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5565..6153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5645..5664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6324..7115,7116..7184))
                     /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     6947..6966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7185..7289)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaacgg gccctctaga ctcgacatga ggttgacgct actttgttgc acctggaggg
      961 aagaacgtat gggagaggaa ggaagcgagt tgcctgtgtg tgcgagctgt ggccagagga
     1021 tctatgacgg ccagtacctc caggccctga atgctgactg gcatgcagac tgcttcaggt
     1081 gctgcgagtg tagcgtctcc ctttcacacc agtactacga gaaggatgga cagctcttct
     1141 gcaagaagga ctactgggcc cgctatggcg agtcttgcca cgggtgctcc gagcacatca
     1201 ccaaagggct ggtcatggtg gctggggagc tgaagtacca ccctgagtgt ttcatctgcc
     1261 tcgcctgtgg aaacttcatt ggcgatgggg acacctacac actggtggag cactccaagc
     1321 tgtactgtgg ccagtgctac taccagaccg tggtaactcc agtcatcgaa cagatcctac
     1381 ctgactcacc cggctcccac ctgccccaca cagtcaccct cgtgtctatc ccagcctctg
     1441 cccatggcaa acgaggcctg tctgtctcca tcgacccccc tcacggccca ccaggatgtg
     1501 gcacggagca ttcgcacact gtccgagtcc aaggagtgga cccaggctgc atgagcccag
     1561 atgtgaagaa ttccatccac gttggagacc ggatcttgga aatcaatggc acgcccatcc
     1621 ggaatgtgcc gctggacgag atcgacttgc tgatccagga gaccagccgc ctgctccagc
     1681 tgaccctcga gcacgacccc catgactcac tgggccatgg gcccgtgtcg gatcccagcc
     1741 ccctgagctc cccggtgcac actcccagcg gacaggcagc cagctctgcc cggcagaaac
     1801 ctgtcttgag gagctgcagt attgacacgt ccccgggtac cagctcacta gcctctccag
     1861 cctcccagcg caaggacctg ggtcgctccg aatccctccg agtggtctgc cgaccacacc
     1921 gaatcttccg gccatctgat ctcatccatg gggaagtgct gggcaagggc tgttttggcc
     1981 aggccatcaa ggtgacacac cgagaaacag gcgaggtgat ggtgatgaag gaacttatcc
     2041 ggtttgatga ggagacccag aggacattcc tcaaggaggt aaaggtcatg cgctgcctgg
     2101 agcaccctaa cgtgctcaag ttcatcggag tgctctacaa ggacaagcgg ctaaacttca
     2161 tcacagagta catcaagggc ggcaccctgc ggggcatcat caagaacatg gacagccagt
     2221 acccgtggag tcagagggtc agctttgcca aggacattgc atcagggatg gcctacctcc
     2281 attcgatgaa catcatccac cgagacctca actcccacaa ctgtctggtc cgtgagaaca
     2341 ggaacgtggt ggtggccgac tttgggctgg ctcgactcat gatcgatgaa aagaatcagt
     2401 ctgaagactt acgcagcctt aagaagccgg accgcaagaa acggtacgaa gaagtggtgg
     2461 gcaaccccta ctggatggcg ccagagatga tcaatggtcg tagttatgat gaaaaggtgg
     2521 acgtgttttc ctttggaatt gtcctctgtg agattatcgg gcgtgtgaat gcagaccctg
     2581 actatctgcc ccgcactatg gactttggcc tcaatgtaag gggtttcctg gaccgctact
     2641 gtccaccaaa ctgtccccca agtttcttcc ccatcactgt gcgctgctgt gacctggatc
     2701 ctgaaaagag accctctttt gtaaagctgg aacaatggct agaaacactt cgcatgcact
     2761 tgtctggtca cctgccactg ggcccacaac tggaacaact ggagcggggc ttctgggaga
     2821 cctaccggcg gggcgagagc tcactgcctg cccaccccga ggtccctgac tagtctagag
     2881 ggccaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
     2941 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3001 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3061 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3121 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3181 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3601 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3781 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3961 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4021 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4081 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4141 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4201 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4261 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4321 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4381 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4441 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4501 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4561 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4621 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4681 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4741 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4801 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4861 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4921 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4981 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5041 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5101 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5161 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5221 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5281 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5341 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5401 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5761 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6001 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6241 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
