LOCUS       sequence-319712-        8373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses KCNIP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8373)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8373)
  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_KCNIP1_OE_1
FEATURES             Location/Qualifiers
     source          1..8373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    616..640
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1292..1316)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1318..1359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1401..1989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1454..1474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1872..1883)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2061..2294
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2372..2493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2409..2428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2463..2482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2533..2668
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2688..2707)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2689..2707)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2717..2734)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2717..2733)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2726..2748)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2875..3330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2962..2981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3172..3193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3356..3460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3461..3529,3530..4321)
                     /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(3679..3698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4492..5080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4981..5000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5234..5251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5368..5389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5404..5409,5410..5427,5428..5434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5442..5458
                     /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     5447..5469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5466..5482
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5466..5482
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5501..5521
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5503..5521
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5549..5775
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5776..5956
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6003..6128
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6621..6854
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7039..7083
                     /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        7258..7758
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7677..7695
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7770..8165
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8225..8342
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tttggcgccg gctcgagggg gcccggggtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactgtcgg gatcaacaag tttgtacaaa aaagttggca ccatgggggc cgtcatgggc
      661 accttctcat ctctgcaaac caaacaaagg cgaccctcga aagataagat tgaagatgag
      721 ctggagatga ccatggtttg ccatcggccc gagggactgg agcagctcga ggcccagacc
      781 aacttcacca agagggagct gcaggtcctt tatcgaggct tcaaaaatga gtgccccagt
      841 ggtgtggtca acgaagacac attcaagcag atctatgctc agtttttccc tcatggagat
      901 gccagcacgt atgcccatta cctcttcaat gccttcgaca ccactcagac aggctccgtg
      961 aagttcgagg actttgtaac cgctctgtcg attttattga gaggaactgt ccacgagaaa
     1021 ctaaggtgga catttaattt gtatgacatc aacaaggacg gatacataaa caaagaggag
     1081 atgatggaca ttgtcaaagc catctatgac atgatgggga aatacacata tcctgtgctc
     1141 aaagaggaca ctccaaggca gcatgtggac gtcttcttcc agaaaatgga caaaaataaa
     1201 gatggcatcg taactttaga tgaatttctt gaatcatgtc aggaggacga caacatcatg
     1261 aggtctctcc agctgtttca aaatgtcatg ttgccaactt tcttgtacaa agtggttggt
     1321 aagcctatcc ctaaccctct cctcggtctc gattctacgt agtaatgagc tagcgctaac
     1381 cggtggcgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     1441 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1501 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     1561 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     1621 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     1681 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     1741 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     1801 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     1861 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     1921 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     1981 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     2041 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     2101 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2161 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2221 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2281 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     2341 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     2401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     2521 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2581 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2641 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     2701 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2761 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2821 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     2881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3121 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3181 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3241 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3301 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3421 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3481 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3541 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3601 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3661 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3721 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3781 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3841 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3901 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3961 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4021 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4081 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4141 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4201 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4261 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4321 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4381 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4441 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4501 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4561 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4621 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4681 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4741 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4801 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4861 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4921 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4981 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5041 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5101 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5161 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5221 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5281 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5341 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5401 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5461 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     5521 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     5581 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     5641 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     5701 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     5761 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5821 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5881 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5941 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     6001 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     6061 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     6121 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     6181 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     6241 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     6301 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     6361 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     6421 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     6481 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     6541 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     6601 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     6661 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     6721 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     6781 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     6841 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     6901 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     6961 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     7021 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     7081 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     7141 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     7201 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccttg
     7261 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     7321 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     7381 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     7441 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     7501 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     7561 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     7621 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     7681 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     7741 accgacctct ctccccaggg ggtaccacca tggccaagcc tttgtctcaa gaagaatcca
     7801 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     7861 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     7921 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     7981 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     8041 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     8101 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     8161 agggcctgca gctgcagtag taagaattct agatcttgag acaaatggca gtattcatcc
     8221 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     8281 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     8341 ttcgggttta ttacagggac agcagagatc cac
//
