LOCUS       sequence-322807-        8475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CLIC5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8475)
  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_CLIC5_OE_1
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    615..639
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1393..1417)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1419..1460
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1502..2090
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1555..1575)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1973..1984)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2162..2395
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2473..2594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2564..2583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2634..2769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2789..2808)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2790..2808)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2818..2835)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2818..2834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2827..2849)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2976..3431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3273..3294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3457..3561
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3562..3630,3631..4422)
                     /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(3780..3799)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4593..5181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5082..5101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5335..5352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5469..5490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5505..5510,5511..5528,5529..5535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5543..5559
                     /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     5548..5570
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5567..5583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5567..5583
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5602..5622
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5604..5622
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5650..5876
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5877..6057
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6104..6229
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6722..6955
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7140..7184
                     /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        7359..7859
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7778..7796
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7871..8266
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8326..8443
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttggcgccgg ctcgaggggg cccggggtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      601 aactgtcggg atcaacaagt ttgtacaaaa aagttggcat gacagactcg gcgacagcta
      661 acggggacga cagggacccc gagatcgagc tctttgtgaa ggctggaatc gatggagaaa
      721 gcatcggcaa ctgtcctttc tctcagcgcc tcttcatgat cctctggctg aaaggagtcg
      781 tgttcaatgt caccactgtg gatctgaaaa gaaagccagc tgacctgcac aacctagccc
      841 ccggcacgca cccgcccttc ctgaccttca acggggacgt gaagacagac gtcaataaga
      901 tcgaggagtt cctggaggag accttgaccc ctgaaaagta ccccaaactg gctgcaaaac
      961 accgggaatc caacacagcg ggcatcgaca tcttttccaa gttttctgcc tacatcaaaa
     1021 ataccaagca gcagaacaat gctgctcttg aaagaggcct aaccaaggct ctaaagaaat
     1081 tggatgacta cctgaacacc cctctaccag aggagattga cgccaacact tgtggggaag
     1141 acaaggggtc ccggcgcaag ttcctggatg gggatgagct gaccctggct gactgcaatc
     1201 tgttgcccaa gctccatgtg gtcaagattg tggccaagaa ataccgcaac tatgatatcc
     1261 cggctgagat gacaggcctg tggcggtacc tcaagaacgc ctatgcccgt gatgagttca
     1321 ccaacacctg tgcagctgac agtgagatcg agttggccta cgctgatgtc gccaaacgcc
     1381 tcagccgatc ctgcccaact ttcttgtaca aagtggttgg taagcctatc cctaaccctc
     1441 tcctcggtct cgattctacg tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga
     1501 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1561 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1621 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1681 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1741 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1801 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1861 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     1921 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     1981 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2041 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     2101 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     2161 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     2221 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     2281 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     2341 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     2401 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     2461 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2581 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     2641 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2701 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2761 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     2821 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2881 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2941 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3181 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3301 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3421 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3481 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3541 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3601 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3661 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3721 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3781 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3841 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3901 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3961 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4021 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4081 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4141 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4201 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4261 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4321 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4381 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5281 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5341 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5401 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5461 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     5521 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5581 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     5641 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     5701 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     5761 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     5821 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     5881 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5941 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6001 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     6061 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     6121 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     6181 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     6241 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     6301 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     6361 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     6421 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     6481 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     6541 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     6601 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     6661 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     6721 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     6781 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     6841 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     6901 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     6961 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     7021 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     7081 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     7141 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     7201 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     7261 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     7321 tatcgtttca gacccacctc ccaaccccga ggggaccctt gcgccttttc caaggcagcc
     7381 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
     7441 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
     7501 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
     7561 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
     7621 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
     7681 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
     7741 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
     7801 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
     7861 gggtaccacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     7921 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     7981 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     8041 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     8101 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     8161 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     8221 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc agctgcagta
     8281 gtaagaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     8341 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     8401 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     8461 cagcagagat ccact
//
