LOCUS       sequence-319339-        8667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CACNG3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8667)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8667)
  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_CACNG3_OE_1
FEATURES             Location/Qualifiers
     source          1..8667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    595..619
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1565..1589)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1591..1632
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1674..2262
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1727..1747)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2145..2156)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2334..2567
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2645..2766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2682..2701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2736..2755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2806..2941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2961..2980)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2962..2980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2990..3007)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2990..3006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2999..3021)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3148..3603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3235..3254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3445..3466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3629..3733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3734..3802,3803..4594)
                     /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(3952..3971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4765..5353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5254..5273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5507..5524
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5641..5662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5677..5682,5683..5700,5701..5707)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5715..5731
                     /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     5720..5742
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5739..5755
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5739..5755
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5774..5794
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5776..5794
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5822..6048
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6049..6229
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6276..6401
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6894..7127
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7312..7356
                     /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        7531..8031
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7950..7968
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             8043..8438
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8498..8615
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgtacaaaa aagttggcat gaggatgtgt gacagaggta tccagatgtt gatcaccact
      661 gtaggagcct ttgccgcttt tagtttaatg accattgcag tgggcacgga ctactggtta
      721 tattccagag gtgtgtgcag gactaaatct acaagtgata atgaaaccag caggaagaat
      781 gaagaagtaa tgacccattc ggggctgtgg aggacctgct gcctagaagg ggctttccga
      841 ggcgtgtgca agaaaatcga tcacttccct gaagatgctg actacgaaca ggacacagcc
      901 gaatatctcc tgcgagctgt gagggcctcc agtgtcttcc ccatcctcag tgtcacgctg
      961 ctgttcttcg gcgggctctg cgtggcagcc agtgagttcc accgcagcag acacaacgtc
     1021 attctcagcg cgggcatctt ttttgtctct gcagggttaa gcaacatcat tggcatcata
     1081 gtttatatat cagccaacgc cggagacccc gggcagcgtg actccaaaaa aagttactcc
     1141 tatggttggt ccttttattt cggagccttc tctttcatca tcgcagaaat tgtaggagtg
     1201 gttgccgtgc acatctatat tgaaaaacat cagcagttac gagccaaatc ccactcggag
     1261 ttcctgaaga aatctacttt tgcccgcctc ccaccctaca ggtatcgatt ccggaggcgg
     1321 tcaagttctc gctccaccga gcccagatcc cgagacctgt cccccatcag caaaggcttc
     1381 cacaccatcc cttccactga catctcgatg ttcaccctct cccgggaccc ctcaaagatc
     1441 accatgggga ccctcctcaa ctccgaccgg gaccacgctt ttctacagtt ccacaattcc
     1501 acacccaaag agttcaaaga gtcactgcat aataatccgg ccaacaggcg caccacgccc
     1561 gtctgcccaa ctttcttgta caaagtggtt ggtaagccta tccctaaccc tctcctcggt
     1621 ctcgattcta cgtagtaatg agctagcgct aaccggtggc gcgttaagtc gacaatcaac
     1681 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1741 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1801 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1861 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1921 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     1981 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2041 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     2101 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2161 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2221 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat
     2281 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     2341 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     2401 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     2461 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     2521 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag
     2581 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     2641 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2701 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2761 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     2821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2941 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     3001 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3061 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3121 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3181 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3241 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3301 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3361 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3421 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3481 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3541 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3601 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5401 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5461 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5521 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5581 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5641 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5701 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     5761 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
     5821 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
     5881 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
     5941 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
     6001 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
     6061 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6121 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6181 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     6241 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     6301 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     6361 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     6421 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     6481 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     6541 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     6601 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     6661 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     6721 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     6781 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     6841 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     6901 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     6961 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     7021 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     7081 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     7141 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     7201 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     7261 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     7321 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     7381 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     7441 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     7501 cagacccacc tcccaacccc gaggggaccc ttgcgccttt tccaaggcag ccctgggttt
     7561 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
     7621 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
     7681 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
     7741 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
     7801 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
     7861 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
     7921 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
     7981 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gggggtacca
     8041 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     8101 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     8161 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     8221 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     8281 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     8341 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     8401 ttcgtgaatt gctgccctct ggttatgtgt gggagggcct gcagctgcag tagtaagaat
     8461 tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     8521 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     8581 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     8641 atccactttg gcgccggctc gaggggg
//
