LOCUS       sequence-334570-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CACNG6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8511)
  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_CACNG6_OE_1
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..610
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1400..1424)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1426..1467
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1509..2097
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1562..1582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1980..1991)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2169..2402
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2480..2601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2517..2536)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2571..2590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2641..2776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2796..2815)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2797..2815)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2825..2842)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2825..2841)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2834..2856)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2983..3438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3070..3089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3280..3301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3464..3568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3569..3637,3638..4429)
                     /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(3787..3806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4600..5188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5089..5108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5342..5359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5476..5497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5512..5517,5518..5535,5536..5542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5550..5566
                     /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     5555..5577
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5574..5590
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5574..5590
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5609..5629
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5611..5629
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5657..5883
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5884..6064
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6111..6236
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6729..6962
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7147..7191
                     /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        7366..7866
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7785..7803
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7878..8273
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8333..8450
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctctctggc taactgtcgg gatcaacaag tttgtacaaa
      601 aaagcaggct ccaccatgat gtggtccaac ttcttcctgc aagaggagaa ccggcggcgg
      661 ggggccgcgg gccggcggcg ggcgcacggg cagggcaggt cggggctgac gcccgagcgc
      721 gaggggaagg tgaagctggc gctgctgctg gccgccgtgg gcgccacgct ggcggtgctg
      781 tccgtgggca ccgagttctg ggtggagctc aacacctaca aggccaacgg cagcgccgtg
      841 tgcgaagcgg cccacctggg gctgtggaag gcgtgcacca agcggctgtg gcaggcggac
      901 gtgcccgtgg acagggacac ctgcggcccc gcggagctgc ccggagaagc aaactgcacc
      961 tattttaaat tcttcaccac gggggagaat gcacgcatct ttcagagaac cacaaagaaa
     1021 gaggtgaatc tggcagctgc ggtgatagca gtgctgggcc tggcagtcat ggccttgggg
     1081 tgcctctgta tcatcatggt gctcagtaaa ggtgcagagt tcctgctccg agttggagcc
     1141 gtctgctttg gcctctcagg cctgctgctc ttggtgagcc tggaggtgtt ccggcattcc
     1201 gtgagggccc tgctgcagag agtcagcccg gagcctcccc cggccccacg cctcacctac
     1261 gagtactcct ggtccctggg ctgcggcgtg ggggccggcc tgatcctgct gttgggggcc
     1321 ggctgctttc tgctgctcac actgccttcc tggccctggg ggtccctctg tcccaagcgg
     1381 gggcaccggg ccaccttgga cccagctttc ttgtacaaag tggttggtaa gcctatccct
     1441 aaccctctcc tcggtctcga ttctacgtag taatgagcta gcgctaaccg gtggcgcgtt
     1501 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1561 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1621 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     1681 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     1741 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     1801 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1861 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     1921 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     1981 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2041 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     2101 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2161 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     2221 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     2281 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     2341 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     2401 cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     2461 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     2521 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2581 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     2641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2701 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2761 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     2821 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2881 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2941 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3001 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3061 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3121 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3181 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3241 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3301 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3361 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3421 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3481 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3541 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3601 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3661 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3721 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3781 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3841 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3901 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3961 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4021 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4081 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4141 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4201 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4261 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4321 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4381 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4441 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4501 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4561 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4621 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4681 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4741 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4801 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4861 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4921 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4981 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5041 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5101 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5161 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5221 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5281 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5341 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5401 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5461 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5521 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5581 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     5641 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     5701 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     5761 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     5821 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     5881 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5941 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6001 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6061 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     6121 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     6181 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     6241 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     6301 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     6361 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     6421 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     6481 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     6541 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     6601 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     6661 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     6721 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     6781 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     6841 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     6901 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     6961 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7021 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7081 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7141 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7201 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7261 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7321 tcaccattat cgtttcagac ccacctccca accccgaggg gacccttgcg ccttttccaa
     7381 ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc
     7441 ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc
     7501 gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt
     7561 tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc
     7621 ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc
     7681 aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag
     7741 gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc
     7801 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
     7861 ccccaggggg taccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     7921 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     7981 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     8041 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     8101 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     8161 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     8221 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcctgcagc
     8281 tgcagtagta agaattctag atcttgagac aaatggcagt attcatccac aattttaaaa
     8341 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     8401 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     8461 acagggacag cagagatcca ctttggcgcc ggctcgaggg ggcccggggt t
//
