LOCUS       sequence-207748-        7913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of murine cyclic GMP-dependent kinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7913)
  AUTHORS   He B, Weber GF
  TITLE     Synergistic activation of the CMV promoter by NF-kappaB P50 and PKG.
  JOURNAL   Biochem Biophys Res Commun. 2004 Aug 13;321(1):13-20. doi: 
            10.1016/j.bbrc.2004.06.101.
  PUBMED    15358208
REFERENCE   2  (bases 1 to 7913)
  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: 
            "10.1016/j.bbrc.2004.06"; journalName: "Biochem Biophys Res Commun";
            date: "2004-08-13- 13"; volume: "321"; issue: "1"; pages: "13-20"
COMMENT     Sequence Label: pEGF/HisB-cGK1
FEATURES             Location/Qualifiers
     source          1..7913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1256..1273
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1277..1309
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1313..1321,1322..1336)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(3519..3536)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3525..3749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3795..4223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3882..3901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4092..4113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4232..4252)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4237..4566
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4417..4552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4479..4498
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4614..4661
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4680..5078
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5236..5357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5273..5292)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5327..5346
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5406..5422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5406..5422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5419..5441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5430..5446
                     /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(5454..5460,5461..5478,5479..5484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5499..5520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5637..5654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5808..6396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5888..5907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6567..7358,7359..7427))
                     /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     7190..7209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7428..7532)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7607..7626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt accatggggg gttctcatca
     1261 tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta
     1321 cgacgatgac gataaggtac ctaaggatcc actagtccag tgtggtggaa ttcggcttag
     1381 catgggcacc ctgcgggatt tacagtatgc gctccaggag aagatcgagg agctgaggca
     1441 gcgggatgct ctcatcgatg agctggagct ggagttggat cagaaggacg aactgatcca
     1501 gaagctgcag aacgaactgg acaagtatcg ctcggtgatc cggccggcca cccagcaggc
     1561 gcagaagcag agcgccagca ccttgcaggg tgaaccgcgc accaagcgcc aggcgatctc
     1621 cgcggagccc accgccttcg acatccagga tctcagccac gtgaccctgc ccttctaccc
     1681 caagagccca cagtcgaagg atctcataaa ggaggctatc cttgacaatg acttcatgaa
     1741 gaacttggag ctgtcacaga tccaggagat tgtggactgt atgtaccccg tggaatacgg
     1801 caaggacagc tgcatcatca aggaaggcga tgtggggtca ctggtgtacg tcatggaaga
     1861 tgggaaggtt gaggtcacaa aagaaggcgt gaagctctgc accatgggtc ctggaaaagt
     1921 gttcggggag ctggctatac tttacaactg tacccggaca gcgaccgtca agactcttgt
     1981 aaatgtgaaa ctctgggcca tcgatcgaca atgttttcaa acaataatga tgaggacagg
     2041 actcatcaag cataccgagt acatggaatt tttaaaaagt gttccaacat tccagagcct
     2101 tcctgatgaa atcctcagca agctggctga tgtcctcgaa gagacccact atgaaaatgg
     2161 agaatatatc atcaggcagg gtgcaagagg agacaccttc ttcatcatca gtaaagggca
     2221 ggtgaatgtt actcgagaag actcaccaag tgaagaccca gtcttcctta gaactttagg
     2281 gaagggagat tggtttggag agaaagcgtt gcagggggag gatgtgagaa cagcaaatgt
     2341 tattgccgca gaagctgtca cctgccttgt gattgaccga gactctttca agcatttgat
     2401 tggaggactg gacgatgttt ctaacaaagc atatgaggac gcagaagcaa aagcaaaata
     2461 tgaagccgaa gctgccttct tcgccaacct gaagctgtct gatttcaaca tcattgacac
     2521 ccttggagtt ggaggtttcg gacgagtaga gctggtccag ttgaaaagtg aagaatccaa
     2581 aacatttgca atgaagatcc tcaagaaacg ccacattgtg gacaccagac agcaggagca
     2641 catccgctca gagaagcaga tcatgcaggg ggctcattct gacttcattg tgaggctgta
     2701 caggacattt aaagacagca aatacttgta tatgttgatg gaagcgtgcc tgggtggaga
     2761 gctctggact attctcaggg ataggggttc gtttgaagac tcaacaacca ggttttacac
     2821 tgcatgtgtg gtagaagcat tcgcctatct gcattccaaa ggaatcattt acagggacct
     2881 caagccggag aatctcatcc tagatcatcg aggctatgcc aaactggttg actttggctt
     2941 tgcaaagaaa ataggatttg gaaagaaaac atggactttt tgtgggactc cagaatatgt
     3001 agccccagag atcatcctga acaaaggcca tgacatttca gctgactact ggtcactagg
     3061 aattctgatg tatgagcttc tgactggcag cccacctttc tcaggcccag atccaatgaa
     3121 aacctacaat atcatactgc gggggattga catgatagaa tttccaaaga agattgcaaa
     3181 aaatgctgct aatttaatta aaaaactatg cagggacaac ccatcagaga ggttaggaaa
     3241 tttgaaaaac ggagtcaaag acattcagaa acacaagtgg tttgagggct ttaattggga
     3301 aggcttaaga aaaggcacct tgacacctcc cataattcca agtgttgcgt cacccacaga
     3361 cacaagcaat tttgacagtt tccctgagga cagcgatgag ccaccacctg atgacaactc
     3421 aggctgggac atagacttct aataagccga attctgcaga tatccagcac agtggcggcc
     3481 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3541 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3601 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3661 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3721 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3781 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3841 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3901 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3961 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4021 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4081 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4141 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4201 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4321 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4381 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4441 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4501 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4561 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     4621 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     4681 tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca
     4741 acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc
     4801 gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg
     4861 tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa
     4921 atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc
     4981 atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc
     5041 gtgaattgct gccctctggt tatgtgtggg agggctaagc acttcgtggc cgaggagcag
     5101 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5161 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5221 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5281 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5341 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5401 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5461 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5521 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5581 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5641 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5701 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5761 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5821 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5881 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5941 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6001 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6061 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6121 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6181 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6241 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6301 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6361 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7501 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7561 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7621 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7681 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7741 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     7801 attgattatt gactaggctt ttgcaaaaag ctttgcaaag atggataaag ttttaaacag
     7861 agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga att
//
