LOCUS       sequence-319713-        8550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CACNG5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8550)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8550)
  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_CACNG5_OE_1
FEATURES             Location/Qualifiers
     source          1..8550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    593..617
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1446..1470)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1472..1513
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1555..2143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1608..1628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2026..2037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2215..2448
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2526..2647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2563..2582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2617..2636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2687..2822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2842..2861)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2843..2861)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2871..2888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2871..2887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2880..2902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3029..3484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3116..3135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3326..3347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3510..3614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3615..3683,3684..4475)
                     /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(3833..3852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4646..5234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5135..5154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5388..5405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5522..5543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5558..5563,5564..5581,5582..5588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5596..5612
                     /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     5601..5623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5620..5636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5620..5636
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5655..5675
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5657..5675
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5703..5929
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5930..6110
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6157..6282
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6775..7008
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7193..7237
                     /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        7412..7912
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7831..7849
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7924..8319
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8379..8496
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cggggttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ctctggctaa ctgtcgggat caacaagttt
      601 gtacaaaaaa gttggcacca tgagtgcctg cgggaggaag gccctgaccc tgctgagcag
      661 tgtctttgct gtctgtggct tgggcctcct gggtatcgcg gtcagcaccg actactggct
      721 gtacctggag gagggtgtga ttgtgcccca gaaccagagc accgagatca agatgtccct
      781 gcactcaggc ctctggcggg tctgcttcct tgcaggtgag gagcgggggc gttgcttcac
      841 catagaatat gtgatgccca tgaacaccca gctgacatcc gagtccacgg tcaatgttct
      901 aaagatgatc cgctcagcca caccattccc tctggtcagc ctcttcttca tgttcattgg
      961 gtttatcctg aacaacatcg gacacatccg tccccaccgg acgatactgg cctttgtctc
     1021 tggcatcttc tttatcctct caggcctctc tctcgtggtg ggcctggtgc tctacatctc
     1081 cagcatcaac gatgagatgc tcaacaggac caaggatgca gagacctact tcaactacaa
     1141 gtatgggtgg tcgtttgcct tcgccgccat ctccttcctt ttaacggaga gtgccggggt
     1201 gatgtctgtg tacctgttta tgaagcggta caccgcggag gacatgtaca ggccccaccc
     1261 tggcttctac cgccctcggc tgagcaactg ctccgattac tcaggccagt tcctacaccc
     1321 agacgcctgg gtcaggggcc gcagcccctc cgacatctcc agcgaggcct ccctgcagat
     1381 gaacagcaac taccccgcct tgctcaagtg ccccgactat gatcagatgt cctcttcacc
     1441 ctgcttgcca actttcttgt acaaagtggt tggtaagcct atccctaacc ctctcctcgg
     1501 tctcgattct acgtagtaat gagctagcgc taaccggtgg cgcgttaagt cgacaatcaa
     1561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     1801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     1861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1921 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     1981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     2161 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     2221 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     2281 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2341 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2401 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     2461 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     2521 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2581 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2641 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     2701 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2761 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2821 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     2881 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2941 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3001 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     3061 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3121 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3181 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3241 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3301 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3361 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3421 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3481 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5341 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5401 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5461 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5521 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5581 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5641 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     5701 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     5761 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
     5821 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     5881 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     5941 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6001 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6061 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     6121 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     6181 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     6241 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     6301 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     6361 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     6421 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     6481 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     6541 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     6601 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     6661 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     6721 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     6781 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     6841 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     6901 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     6961 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     7021 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     7081 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     7141 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     7201 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     7261 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     7321 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     7381 tcagacccac ctcccaaccc cgaggggacc cttgcgcctt ttccaaggca gccctgggtt
     7441 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
     7501 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
     7561 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
     7621 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
     7681 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
     7741 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     7801 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
     7861 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggtacc
     7921 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     7981 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     8041 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     8101 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     8161 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     8221 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     8281 attcgtgaat tgctgccctc tggttatgtg tgggagggcc tgcagctgca gtagtaagaa
     8341 ttctagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     8401 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     8461 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     8521 gatccacttt ggcgccggct cgagggggcc
//
