LOCUS       sequence-318726-        8490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses KCNG4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8490)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8490)
  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"
COMMENT     Sequence Label: IDG_KCNG4_OE_1
FEATURES             Location/Qualifiers
     source          1..8490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    609..633
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1402..1426)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1428..1469
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1511..2099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1564..1584)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1982..1993)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2171..2404
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2482..2603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2519..2538)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2573..2592
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2643..2778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2798..2817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2799..2817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2827..2844)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2827..2843)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2836..2858)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2985..3440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3072..3091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3282..3303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3466..3570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3571..3639,3640..4431)
                     /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     complement(3789..3808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4602..5190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5091..5110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5344..5361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5478..5499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5514..5519,5520..5537,5538..5544)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5552..5568
                     /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)."
     primer_bind     5557..5579
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5576..5592
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5576..5592
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5611..5631
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5613..5631
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5659..5885
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5886..6066
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6113..6238
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6731..6964
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7149..7193
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7368..7868
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7787..7805
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7880..8275
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8335..8452
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccggctcgag ggggcccggg gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactgt
      601 cgggatcaac aagtttgtac aaaaaagttg gcatgcccat gccttccaga gacgggggcc
      661 tgcatcccag acaccaccac tatggttccc acagcccttg gagtcagctc ctgtccagcc
      721 ccatggagac gccgtccatc aagggccttt actaccggag ggtgcggaag gtgggtgccc
      781 tggacgcctc cccagtggac ctgaagaagg agatcctgat caacgtgggg ggcaggaggt
      841 atctcctccc ctggagcaca ctggaccggt tcccgctgag ccgcctgagc aaactcaggc
      901 tctgtcggag ctacgaggag atcgtgcagc tctgcgatga ttacgacgag gacagccagg
      961 agttcttctt cgacaggagc cccagcgcct tcggggtgat cgtgagcttc ctggcggccg
     1021 ggaagctggt gcttctgcag gagatgtgcg cgctgtcctt ccaggaggag ctggcctact
     1081 ggggcatcga ggaggcccac ctggagaggt gctgcctgcg gaagctgctg aggaagctgg
     1141 aggagctgga ggagctggcc aagctgcaca gggaggacgt actgaggcag cagagggaga
     1201 cccgccgccc cgcctcgcac tcctcgcgct ggggcctgtg catgaaccgg ctgcgcgaga
     1261 tggtggaaaa cccgcagtcc gggctgcccg ggaaggtctt cgcttgcctc tccatcctct
     1321 tcgtggccac cacagccgtc agcctgtgtg tcagcaccat gcccgacctc agggcagagg
     1381 aggaccaggt gagcggcctc tgcccaactt tcttgtacaa agtggttggt aagcctatcc
     1441 ctaaccctct cctcggtctc gattctacgt agtaatgagc tagcgctaac cggtggcgcg
     1501 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1561 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1621 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1681 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1741 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1801 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1861 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     1921 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     1981 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2041 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     2101 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2161 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     2221 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     2281 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2341 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2401 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2461 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     2521 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2581 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     2641 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2701 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2761 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     2821 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2881 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2941 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3421 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3481 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3541 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3601 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3661 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3721 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3781 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3841 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3901 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3961 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4021 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4081 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4141 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4201 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4261 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4321 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4381 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4441 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5281 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5341 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5401 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5461 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5521 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5581 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     5641 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     5701 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     5761 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     5821 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     5881 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5941 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6001 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6061 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     6121 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     6181 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     6241 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     6301 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     6361 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     6421 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     6481 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     6541 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     6601 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     6661 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     6721 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     6781 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     6841 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     6901 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     6961 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     7021 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     7081 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     7141 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     7201 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     7261 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     7321 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccttg cgccttttcc
     7381 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     7441 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     7501 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     7561 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     7621 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     7681 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     7741 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     7801 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     7861 ctccccaggg ggtaccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     7921 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     7981 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     8041 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     8101 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     8161 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     8221 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggcctgca
     8281 gctgcagtag taagaattct agatcttgag acaaatggca gtattcatcc acaattttaa
     8341 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     8401 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     8461 ttacagggac agcagagatc cactttggcg
//
