LOCUS       sequence-319633-        8703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CACNG4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8703)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8703)
  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_CACNG4_OE_1
FEATURES             Location/Qualifiers
     source          1..8703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    531..555
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1537..1561)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1563..1604
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1646..2234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1699..1719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2117..2128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2306..2539
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2617..2738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2654..2673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2708..2727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2778..2913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2933..2952)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2934..2952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2962..2979)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2962..2978)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2971..2993)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3120..3575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3207..3226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3417..3438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3601..3705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3706..3774,3775..4566)
                     /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(3924..3943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4737..5325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5226..5245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5479..5496
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5613..5634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5649..5654,5655..5672,5673..5679)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5687..5703
                     /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     5692..5714
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5711..5727
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5711..5727
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5746..5766
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5748..5766
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5794..6020
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6021..6201
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6248..6373
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6866..7099
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7284..7328
                     /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        7503..8003
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7922..7940
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             8015..8410
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8470..8587
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact gtcgggatca acaagtttgt
      541 acaaaaaagt tggcatggtg cgatgcgacc gcgggctgca gatgctgctg accacggccg
      601 gagccttcgc cgccttctcg ctcatggcca tcgccatcgg caccgactac tggctgtact
      661 ccagcgcgca catctgcaac ggcaccaacc tgaccatgga cgacgggccc ccgccccgcc
      721 gcgcccgcgg cgacctcacc cactctggtc tgtggcgggt gtgctgcatc gaagggatct
      781 ataaagggca ctgcttccgg atcaatcact tcccagagga caatgactac gaccacgaca
      841 gctcggagta cctcctccgc atcgtgcgag cctccagcgt cttccccatc ctcagcacca
      901 tcctgctcct gctgggtggc ctgtgcatcg gtgctggcag gatctacagc cgcaagaaca
      961 acatcgtcct cagtgccggc atcctcttcg tggctgcagg cctcagtaac atcatcggta
     1021 tcatcgtcta catttccagc aacacaggtg acccgagtga caagcgggac gaagacaaaa
     1081 agaaccatta caactacggc tggtcttttt actttggagc tctgtctttc attgtggctg
     1141 agaccgtggg cgtcctggct gtaaacattt acattgagaa aaataaagag ttgaggttta
     1201 agaccaaacg ggaattcctt aaggcgtctt cctcttctcc ttatgccagg atgccgagct
     1261 acaggtaccg gcgacggcgc tcgaggtcca gctcaaggtc cactgaggcc tcgccctcca
     1321 gggacgtgtc gcccatgggc ctgaagatca caggggccat ccccatgggg gagctgtcca
     1381 tgtacacgct gtccagggag cccctcaagg tgaccaccgc agccagctac agccccgacc
     1441 aggaggccag cttcctgcag gtgcatgact ttttccagca ggacctgaag gaaggtttcc
     1501 acgtcagcat gctgaaccga cggacgaccc ctgtgtgccc aactttcttg tacaaagtgg
     1561 ttggtaagcc tatccctaac cctctcctcg gtctcgattc tacgtagtaa tgagctagcg
     1621 ctaaccggtg gcgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     1681 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1741 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1801 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     1861 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     1921 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     1981 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2041 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2101 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2161 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2221 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     2281 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     2341 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2521 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     2581 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     2641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2701 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     2761 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2821 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2881 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     2941 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     3001 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     3061 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     3121 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3181 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3241 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3301 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3361 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3421 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3481 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3541 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3601 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3661 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3721 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3781 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3841 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3901 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3961 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4021 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4081 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4141 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4201 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4261 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4321 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4381 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4441 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4501 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4561 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4621 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4681 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4741 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4801 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4861 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4921 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4981 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5041 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5101 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5161 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5221 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5281 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5341 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5401 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5461 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5521 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5581 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5641 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5701 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     5761 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     5821 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     5881 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     5941 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     6001 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     6061 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6121 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6181 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     6241 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     6301 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     6361 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     6421 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     6481 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     6541 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     6601 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     6661 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     6721 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     6781 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     6841 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     6901 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     6961 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     7021 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     7081 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     7141 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     7201 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     7261 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     7321 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     7381 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     7441 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     7501 ccttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
     7561 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
     7621 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
     7681 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
     7741 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
     7801 cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg
     7861 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
     7921 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
     7981 gaatcaccga cctctctccc cagggggtac caccatggcc aagcctttgt ctcaagaaga
     8041 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     8101 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     8161 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     8221 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     8281 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     8341 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     8401 gtgggagggc ctgcagctgc agtagtaaga attctagatc ttgagacaaa tggcagtatt
     8461 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     8521 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     8581 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagggggc
     8641 ccggggttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     8701 ccg
//
