LOCUS       sequence-319637-        8319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CATSPER2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8319)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8319)
  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_CATSPER2_OE_1
FEATURES             Location/Qualifiers
     source          1..8319
                     /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(1153..1177)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1179..1220
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1262..1850
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1315..1335)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1733..1744)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1922..2155
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2233..2354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2324..2343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2394..2529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2549..2568)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2550..2568)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2578..2595)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2578..2594)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2587..2609)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2736..3191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2823..2842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3033..3054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3217..3321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3322..3390,3391..4182)
                     /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(3540..3559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4353..4941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4842..4861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5095..5112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5229..5250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5265..5270,5271..5288,5289..5295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5303..5319
                     /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     5308..5330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5327..5343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5327..5343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5362..5382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5364..5382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5410..5636
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5637..5817
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5864..5989
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6482..6715
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6900..6944
                     /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        7119..7619
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7538..7556
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7631..8026
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8086..8203
                     /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 tggcatggcc gcttaccaac aagaagagca gatgcagctt ccccgagctg
      601 atgccattcg ttcacgtctc atcgatactt tctctctcat tgagcatttg caaggcttga
      661 gccaagctgt gccgcggcac actatcagga agttacttga tccttcccgc cagaagaaac
      721 ttgtattggg agatcaacac cagctagtgc gtttctctat aaagcctcag cgtatagaac
      781 agatttcaca tgcccagagg ctgttgagca ggcttcatgt gcgctgcagt cagaggccac
      841 ctctttcttt gtgggccgga tgggtccttg agtgtcctct cttcaaaaac ttcatcatct
      901 tcctggtctt tttgaatacg atcatattga tggttgaaat agaattgctg gaatccacaa
      961 ataccaaact atggccattg aagctgacct tggaggtggc agcttggttt atcttgctta
     1021 ttttcatcct ggagatcctt cttaagtggc tatccaactt ttctgttttc tggaagagtg
     1081 cctggaatgt ctttgacttt gttgttacca tgttggtaag gatagagatc ctgagggttc
     1141 gtttagtggg atgcccaact ttcttgtaca aagtggttgg taagcctatc cctaaccctc
     1201 tcctcggtct cgattctacg tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga
     1261 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1321 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1381 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1441 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1501 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1561 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1621 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     1681 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     1741 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     1801 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     1861 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     1921 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     1981 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     2041 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     2101 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     2161 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     2221 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2341 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     2401 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2461 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2521 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     2581 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2641 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2701 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     2761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3181 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3421 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3481 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3541 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3601 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3661 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3721 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3781 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3841 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3901 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3961 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4021 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4081 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4141 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5041 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5101 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5161 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5221 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     5281 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5341 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     5401 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     5461 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     5521 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     5581 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     5641 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5701 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5761 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     5821 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     5881 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     5941 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     6001 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     6061 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     6121 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     6181 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     6241 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     6301 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     6361 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     6421 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     6481 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     6541 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     6601 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     6661 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     6721 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     6781 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     6841 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     6901 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     6961 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     7021 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     7081 tatcgtttca gacccacctc ccaaccccga ggggaccctt gcgccttttc caaggcagcc
     7141 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
     7201 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
     7261 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
     7321 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
     7381 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
     7441 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
     7501 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
     7561 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
     7621 gggtaccacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     7681 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     7741 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     7801 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     7861 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     7921 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     7981 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc agctgcagta
     8041 gtaagaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     8101 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     8161 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     8221 cagcagagat ccactttggc gccggctcga gggggcccgg ggttattaat agtaatcaat
     8281 tacggggtca ttagttcata gcccatatat ggagttccg
//
