LOCUS       sequence-319638-        8430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses CLIC3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8430)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8430)
  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_CLIC3_OE_1
FEATURES             Location/Qualifiers
     source          1..8430
                     /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(1264..1288)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1290..1331
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1373..1961
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1426..1446)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1844..1855)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2033..2266
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2344..2465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2381..2400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2435..2454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2505..2640
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2660..2679)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2661..2679)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2689..2706)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2689..2705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2698..2720)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2847..3302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2934..2953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3144..3165
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3328..3432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3433..3501,3502..4293)
                     /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(3651..3670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4464..5052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4953..4972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5206..5223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5340..5361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5376..5381,5382..5399,5400..5406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5414..5430
                     /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     5419..5441
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5438..5454
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5438..5454
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5473..5493
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5475..5493
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5521..5747
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5748..5928
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5975..6100
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6593..6826
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7011..7055
                     /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        7230..7730
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7649..7667
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7742..8137
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8197..8314
                     /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 tggcatggcg gagaccaagc tccagctgtt tgtcaaggcg agtgaggacg
      601 gggagagcgt gggtcactgc ccctcctgcc agcggctctt catggtcctg ctcctcaagg
      661 gcgtaccttt caccctcacc acggtggaca cgcgcaggtc cccggacgtg ctgaaggact
      721 tcgcccccgg ctcgcagctg cccatcctgc tctatgacag cgacgccaag acagacacgc
      781 tgcagatcga ggactttctg gaggagacgc tggggccgcc cgacttcccc agcctggcgc
      841 ctcgttacag ggagtccaac accgccggca acgacgtttt ccacaagttc tccgcgttca
      901 tcaagaaccc ggtgcccgcg caggacgaag ccctgtacca gcagctgctg cgcgccctcg
      961 ccaggctgga cagctacctg cgcgcgcccc tggagcacga gctggcgggg gagccgcagc
     1021 tgcgcgagtc ccgccgccgc ttcctggacg gcgacaggct cacgctggcc gactgcagcc
     1081 tcctgcccaa gctgcacatc gtcgacacgg tgtgcgcgca cttccgccag gcgcccatcc
     1141 ccgcggagct gcgcggcgta cgccgctacc tggacagcgc gatgcaggag aaagagttca
     1201 aatacacgtg tccgcacagc gccgagatcc tggcggccta ccggcccgcc gtgcaccccc
     1261 gctacccaac tttcttgtac aaagtggttg gtaagcctat ccctaaccct ctcctcggtc
     1321 tcgattctac gtagtaatga gctagcgcta accggtggcg cgttaagtcg acaatcaacc
     1381 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1441 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1501 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1561 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1621 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1681 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1741 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     1801 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1861 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     1921 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
     1981 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     2041 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     2101 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2161 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2221 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt
     2281 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     2341 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2401 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2461 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     2521 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2581 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2641 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     2701 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2761 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2821 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     2881 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2941 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3001 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3061 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3121 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3181 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3241 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     3301 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3421 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3481 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3541 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3601 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3661 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3721 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3781 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3841 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3901 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3961 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4021 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4081 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4141 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4201 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4261 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4321 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4381 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4441 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4501 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4561 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4621 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4681 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4741 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4801 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4861 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4921 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4981 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5041 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5101 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5161 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5221 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5281 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5341 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     5401 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5461 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa
     5521 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
     5581 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
     5641 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
     5701 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
     5761 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5821 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5881 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     5941 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     6001 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     6061 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     6121 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     6181 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     6241 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     6301 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     6361 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     6421 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     6481 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     6541 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     6601 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     6661 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     6721 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     6781 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     6841 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     6901 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     6961 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     7021 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     7081 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     7141 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     7201 agacccacct cccaaccccg aggggaccct tgcgcctttt ccaaggcagc cctgggtttg
     7261 cgcagggacg cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt
     7321 ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat cttcgccgct acccttgtgg
     7381 gccccccggc gacgcttcct gctccgcccc taagtcggga aggttccttg cggttcgcgg
     7441 cgtgccggac gtgacaaacg gaagccgcac gtctcactag taccctcgca gacggacagc
     7501 gccagggagc aatggcagcg cgccgaccgc gatgggctgt ggccaatagc ggctgctcag
     7561 cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg
     7621 tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca
     7681 cgtcggcagt cggctccctc gttgaccgaa tcaccgacct ctctccccag ggggtaccac
     7741 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     7801 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     7861 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     7921 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     7981 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     8041 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     8101 tcgtgaattg ctgccctctg gttatgtgtg ggagggcctg cagctgcagt agtaagaatt
     8161 ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     8221 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     8281 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     8341 tccactttgg cgccggctcg agggggcccg gggttattaa tagtaatcaa ttacggggtc
     8401 attagttcat agcccatata tggagttccg
//
