LOCUS       sequence-207852-        7131 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 7131)
  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 7131)
  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: pCR3.1-cGK1
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2869..2886)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2875..3099
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3231..3819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3311..3330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4148..4195
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4220..4239
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4427..5221)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4427..5221)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4538..4557)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5148..5167
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5256..5613)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5270..5405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5422..5613
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5570..5590
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5673..6464,6465..6533))
                     /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     6296..6315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6534..6638)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6665..7120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6752..6771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6962..6983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcggcttag catgggcacc ctgcgggatt tacagtatgc gctccaggag aagatcgagg
      781 agctgaggca gcgggatgct ctcatcgatg agctggagct ggagttggat cagaaggacg
      841 aactgatcca gaagctgcag aacgaactgg acaagtatcg ctcggtgatc cggccggcca
      901 cccagcaggc gcagaagcag agcgccagca ccttgcaggg tgaaccgcgc accaagcgcc
      961 aggcgatctc cgcggagccc accgccttcg acatccagga tctcagccac gtgaccctgc
     1021 ccttctaccc caagagccca cagtcgaagg atctcataaa ggaggctatc cttgacaatg
     1081 acttcatgaa gaacttggag ctgtcacaga tccaggagat tgtggactgt atgtaccccg
     1141 tggaatacgg caaggacagc tgcatcatca aggaaggcga tgtggggtca ctggtgtacg
     1201 tcatggaaga tgggaaggtt gaggtcacaa aagaaggcgt gaagctctgc accatgggtc
     1261 ctggaaaagt gttcggggag ctggctatac tttacaactg tacccggaca gcgaccgtca
     1321 agactcttgt aaatgtgaaa ctctgggcca tcgatcgaca atgttttcaa acaataatga
     1381 tgaggacagg actcatcaag cataccgagt acatggaatt tttaaaaagt gttccaacat
     1441 tccagagcct tcctgatgaa atcctcagca agctggctga tgtcctcgaa gagacccact
     1501 atgaaaatgg agaatatatc atcaggcagg gtgcaagagg agacaccttc ttcatcatca
     1561 gtaaagggca ggtgaatgtt actcgagaag actcaccaag tgaagaccca gtcttcctta
     1621 gaactttagg gaagggagat tggtttggag agaaagcgtt gcagggggag gatgtgagaa
     1681 cagcaaatgt tattgccgca gaagctgtca cctgccttgt gattgaccga gactctttca
     1741 agcatttgat tggaggactg gacgatgttt ctaacaaagc atatgaggac gcagaagcaa
     1801 aagcaaaata tgaagccgaa gctgccttct tcgccaacct gaagctgtct gatttcaaca
     1861 tcattgacac ccttggagtt ggaggtttcg gacgagtaga gctggtccag ttgaaaagtg
     1921 aagaatccaa aacatttgca atgaagatcc tcaagaaacg ccacattgtg gacaccagac
     1981 agcaggagca catccgctca gagaagcaga tcatgcaggg ggctcattct gacttcattg
     2041 tgaggctgta caggacattt aaagacagca aatacttgta tatgttgatg gaagcgtgcc
     2101 tgggtggaga gctctggact attctcaggg ataggggttc gtttgaagac tcaacaacca
     2161 ggttttacac tgcatgtgtg gtagaagcat tcgcctatct gcattccaaa ggaatcattt
     2221 acagggacct caagccggag aatctcatcc tagatcatcg aggctatgcc aaactggttg
     2281 actttggctt tgcaaagaaa ataggatttg gaaagaaaac atggactttt tgtgggactc
     2341 cagaatatgt agccccagag atcatcctga acaaaggcca tgacatttca gctgactact
     2401 ggtcactagg aattctgatg tatgagcttc tgactggcag cccacctttc tcaggcccag
     2461 atccaatgaa aacctacaat atcatactgc gggggattga catgatagaa tttccaaaga
     2521 agattgcaaa aaatgctgct aatttaatta aaaaactatg cagggacaac ccatcagaga
     2581 ggttaggaaa tttgaaaaac ggagtcaaag acattcagaa acacaagtgg tttgagggct
     2641 ttaattggga aggcttaaga aaaggcacct tgacacctcc cataattcca agtgttgcgt
     2701 cacccacaga cacaagcaat tttgacagtt tccctgagga cagcgatgag ccaccacctg
     2761 atgacaactc aggctgggac atagacttct aataagccga attctgcaga tatccagcac
     2821 agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     2881 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2941 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3001 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3061 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3121 gtggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3961 agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc
     4021 cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat
     4081 tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg
     4141 cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata
     4201 gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg
     4261 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     4321 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
     4381 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
     4441 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     4501 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     4561 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     4621 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     4681 cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     4741 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     4801 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     4861 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     4921 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     4981 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
     5041 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     5101 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     5161 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     5221 tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa
     5281 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     5341 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     5401 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     5461 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     5521 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     5581 cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa taaatcaatc
     5641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6661 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6721 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6781 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6841 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6901 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6961 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7021 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7081 tttaacgcga attttaacaa aatattaacg cttacaattt acgcgcgcgt t
//
