LOCUS       sequence-319654-        8472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses KCNIP4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8472)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8472)
  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_KCNIP4_OE_1
FEATURES             Location/Qualifiers
     source          1..8472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        44..347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        348..551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     502..522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    574..598
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1349..1373)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1375..1416
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1458..2046
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1511..1531)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1929..1940)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2118..2351
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2429..2550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2466..2485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2520..2539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2590..2725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2745..2764)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2746..2764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2774..2791)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2774..2790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2783..2805)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2932..3387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3019..3038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3229..3250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3413..3517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3518..3586,3587..4378)
                     /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(3736..3755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4549..5137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5038..5057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5291..5308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5425..5446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5461..5466,5467..5484,5485..5491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5499..5515
                     /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     5504..5526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5523..5539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5523..5539
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5558..5578
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5560..5578
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5606..5832
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5833..6013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6060..6185
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6678..6911
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7096..7140
                     /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        7315..7815
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7734..7752
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7827..8222
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8282..8399
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
       61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      541 taagcagagc tctctggcta actgtcggga tcaacaagtt tgtacaaaaa agttggcatg
      601 aatgtgggga gggtggaaag catttcggct cagctggagg aggccagctc tacaggcggt
      661 ttcctgtacg ctcagaacag caccaagcgc agcattaaag agcggctcat gaagctcttg
      721 ccctgctcag ctgccaaaac gtcgtctcct gctattcaaa acagcgtgga agatgaactg
      781 gagatggcca ccgtcaggca tcggcccgaa gcccttgagc ttctggaagc ccagagcaaa
      841 tttaccaaga aagagcttca gatcctttac agaggattta agaacgaatg ccccagtggt
      901 gttgttaatg aagaaacctt caaagagatt tactcgcagt tctttccaca gggagactct
      961 acaacatatg cacattttct gttcaatgca tttgatacag accacaatgg agctgtgagt
     1021 ttcgaggatt tcatcaaagg tctttccatt ttgctccggg ggacagtaca agaaaaactc
     1081 aattgggcat ttaatctgta tgacataaat aaagatggct acatcactaa agaggaaatg
     1141 cttgatataa tgaaagcaat atacgatatg atgggtaaat gtacatatcc tgtcctcaaa
     1201 gaagatgctc ccagacaaca cgttgaaaca ttttttcaga aaatggacaa aaataaagat
     1261 ggggttgtta ccatagatga gttcattgaa agctgccaaa aagatgaaaa cataatgcgc
     1321 tccatgcagc tctttgaaaa tgtgatttac ccaactttct tgtacaaagt ggttggtaag
     1381 cctatcccta accctctcct cggtctcgat tctacgtagt aatgagctag cgctaaccgg
     1441 tggcgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1501 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1561 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1621 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1681 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1741 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1801 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     1861 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     1921 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     1981 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2041 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     2101 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     2161 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2221 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     2281 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     2341 aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc
     2401 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     2461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2521 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     2581 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2641 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2701 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     2761 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2821 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2881 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2941 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3001 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3061 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3121 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3181 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3241 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3301 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3361 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5161 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5221 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5281 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5341 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5401 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5461 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5521 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     5581 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
     5641 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
     5701 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
     5761 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
     5821 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5881 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5941 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6001 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     6061 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     6121 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     6181 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     6241 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     6301 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     6361 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     6421 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     6481 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     6541 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     6601 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     6661 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     6721 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     6781 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     6841 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     6901 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     6961 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     7021 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     7081 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     7141 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     7201 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     7261 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccttgcgc
     7321 cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg
     7381 aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc
     7441 cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt
     7501 cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc
     7561 actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg
     7621 gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg
     7681 gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc
     7741 gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc
     7801 gacctctctc cccagggggt accaccatgg ccaagccttt gtctcaagaa gaatccaccc
     7861 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     7921 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     7981 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     8041 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     8101 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     8161 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     8221 gcctgcagct gcagtagtaa gaattctaga tcttgagaca aatggcagta ttcatccaca
     8281 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     8341 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     8401 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg gcccggggtt
     8461 attaatagta at
//
