LOCUS       sequence-11471-e        7748 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 7748)
  AUTHORS   Tessier M, Woodgett JR
  TITLE     Role of the Phox homology domain and phosphorylation in activation 
            of serum and glucocorticoid-regulated kinase-3.
  JOURNAL   J Biol Chem. 2006 Aug 18. 281(33):23978-89.
  PUBMED    16790420
REFERENCE   2  (bases 1 to 7748)
  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. 2006 Aug 18. 281(33):23978-89."
COMMENT     Sequence Label: Flag SGK3 wt - pCMV
FEATURES             Location/Qualifiers
     source          1..7748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2507..3129
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2706..2725)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3153..3173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3171..3468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3522..4316
                     /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(3576..3595)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4186..4205
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4968..5102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5005..5024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5059..5078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5107..5126)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5108..5126)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5140..5156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5140..5156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5153..5175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5164..5180
                     /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(5188..5194,5195..5212,5213..5218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5233..5254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5371..5388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5542..6130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5622..5641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6301..7092,7093..7161))
                     /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     6924..6943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7162..7266)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7293..7721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7380..7399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7590..7611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgat gcaaagagat
     1021 cacaccatgg actacaagga aagctgccca agtgtaagca ttcccagctc cgatgaacac
     1081 agagagaaaa agaagaggtt tactgtttat aaagttctgg tttcagtggg aagaagtgaa
     1141 tggtttgtct tcaggagata tgcagagttt gataaacttt ataacacttt aaaaaaacag
     1201 tttcctgcta tggccctgaa gattcctgcc aagagaatat ttggtgataa ttttgatcca
     1261 gattttatta aacaaagacg agcaggacta aacgaattca ttcagaacct agttaggtat
     1321 ccagaacttt ataaccatcc agatgtcaga gcattccttc aaatggacag tccaaaacac
     1381 cagtcagatc catctgaaga tgaggatgaa agaagttctc agaagctaca ctctacctca
     1441 cagaacatca acctgggacc gtctggaaat cctcatgcca aaccaactga ctttgatttc
     1501 ttaaaagtta ttggaaaagg cagctttggc aaggttcttc ttgcaaaacg gaaactggat
     1561 ggaaaatttt atgctgtcaa agtgttacag aaaaaaatag ttctcaacag aaaagagcaa
     1621 aaacatatta tggctgaacg taatgtgctc ttgaaaaatg tgaaacatcc gtttttggtt
     1681 ggattgcatt attccttcca aacaactgaa aagctttatt ttgttctgga ttttgttaat
     1741 ggaggggagc tttttttcca cttacaaaga gaacggtcct ttcctgagca cagagctagg
     1801 ttttacgctg ctgaaattgc tagtgcattg ggttacttac attccatcaa aatagtatac
     1861 agagacttga aaccagaaaa tattcttttg gattcagtag gacatgttgt cttaacagat
     1921 tttgggcttt gtaaagaagg aattgctatt tctgacacca ctaccacatt ttgtgggaca
     1981 ccagagtatc ttgcacctga agtaattaga aaacagccct atgacaatac tgtagattgg
     2041 tggtgccttg gggctgttct gtatgaaatg ctgtatggat tgcctccttt ttattgccga
     2101 gatgttgctg aaatgtatga caatatcctt cacaaacccc taagtttgag gccaggagtg
     2161 agtcttacag cctggtccat tctggaagaa ctcctagaaa aagacaggca aaatcgactt
     2221 ggtgccaagg aagactttct tgaaattcag aatcatcctt tttttgaatc actcagctgg
     2281 gctgaccttg tacaaaagaa gattccacca ccatttaatc ctaatgtggc tggaccagat
     2341 gatatcagaa actttgacac agcatttaca gaagaaacag ttccatattc tgtgtgtgta
     2401 tcttctgact attctatagt gaatgccagt gtattggagg cagatgatgc attcgttggt
     2461 ttctcttatg cacctccttc agaagactta tttttgtgag gatcccgggt ggcatccctg
     2521 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2581 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2641 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2701 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2761 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     2821 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     2881 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     2941 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3001 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3061 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3121 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3241 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3301 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3361 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3421 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca
     3481 gcttgggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3541 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3601 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3661 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3721 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3781 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3841 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3901 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3961 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4021 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4081 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4141 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4201 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4261 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4321 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4381 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4441 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc
     4501 cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca
     4561 gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga
     4621 actgcaggag tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg
     4681 cctgatacag aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct
     4741 gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc
     4801 cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4861 cgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4921 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5101 tcaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     5161 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5221 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5281 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5521 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5701 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5941 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7261 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     7321 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7381 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7441 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7501 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     7561 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     7621 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     7681 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     7741 tacaattt
//
