LOCUS       sequence-319579-        8463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CLIC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8463)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8463)
  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_CLIC2_OE_1
FEATURES             Location/Qualifiers
     source          1..8463
                     /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(1297..1321)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1323..1364
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1406..1994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1459..1479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1877..1888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2066..2299
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2377..2498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2414..2433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2468..2487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2538..2673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2693..2712)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2694..2712)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2722..2739)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2722..2738)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2731..2753)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2880..3335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2967..2986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3177..3198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3361..3465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3466..3534,3535..4326)
                     /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(3684..3703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4497..5085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4986..5005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5239..5256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5373..5394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5409..5414,5415..5432,5433..5439)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5447..5463
                     /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     5452..5474
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5471..5487
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5471..5487
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5506..5526
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5508..5526
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5554..5780
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5781..5961
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6008..6133
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6626..6859
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7044..7088
                     /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        7263..7763
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7682..7700
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7775..8170
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8230..8347
                     /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 tggcatgtca ggcctgcggc ccggcactca agtggaccct gagattgagc
      601 tttttgtaaa ggctggaagt gatggagaga gtattggaaa ctgtcccttt tgccaacgcc
      661 ttttcatgat cctctggctt aaaggagtta aatttaatgt gacaactgtt gacatgacca
      721 gaaagcctga agaactaaag gacttagccc caggtaccaa tcctccgttc ctggtgtata
      781 acaaggagtt gaaaacagac ttcattaaaa ttgaggagtt tttagaacaa accctggctc
      841 ctccaaggta ccctcacctg agtcccaagt acaaggagtc ttttgatgtg ggctgtaacc
      901 tctttgccaa gttttctgca tacattaaga atacacaaaa ggaggcaaat aagaattttg
      961 aaaaatctct gctcaaagaa ttcaagcgtc tggatgacta cttaaacacc ccacttctgg
     1021 atgaaattga tccagacagt gctgaggaac ccccagtttc cagaagacta ttcttggatg
     1081 gggaccagct aacactggct gattgtagct tgttacccaa gctgaacatt attaaagttg
     1141 ctgccaagaa atatcgtgac tttgacattc cagcagaatt ctcaggagtc tggcgttatc
     1201 tccacaatgc ctatgcccgt gaagaattta cccacacgtg tcctgaagac aaagaaattg
     1261 aaaatactta cgcaaatgtg gctaaacaga agagttaccc aactttcttg tacaaagtgg
     1321 ttggtaagcc tatccctaac cctctcctcg gtctcgattc tacgtagtaa tgagctagcg
     1381 ctaaccggtg gcgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     1441 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1501 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1561 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     1621 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     1681 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     1741 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     1801 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     1861 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     1921 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     1981 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     2041 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     2101 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2161 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2221 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2281 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     2341 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     2401 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2461 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     2521 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2581 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2641 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     2701 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2761 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2821 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     2881 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2941 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3001 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3061 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3241 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3301 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5281 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5341 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5401 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5461 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     5521 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     5581 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     5641 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     5701 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     5761 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     5821 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5881 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5941 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     6001 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     6061 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     6121 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     6181 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     6241 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     6301 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     6361 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     6421 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     6481 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     6541 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     6601 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     6661 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     6721 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     6781 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     6841 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     6901 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     6961 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     7021 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     7081 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     7141 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     7201 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     7261 ccttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
     7321 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
     7381 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
     7441 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
     7501 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
     7561 cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg
     7621 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
     7681 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
     7741 gaatcaccga cctctctccc cagggggtac caccatggcc aagcctttgt ctcaagaaga
     7801 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     7861 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     7921 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     7981 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     8041 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     8101 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     8161 gtgggagggc ctgcagctgc agtagtaaga attctagatc ttgagacaaa tggcagtatt
     8221 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     8281 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     8341 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagggggc
     8401 ccggggttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     8461 ccg
//
