LOCUS       sequence-319636-        8268 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Overexpresses PLLP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8268)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: IDG_PLLP_OE_1
FEATURES             Location/Qualifiers
     source          1..8268
                     /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    complement(1102..1126)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1128..1169
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1211..1799
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1264..1284)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1682..1693)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1871..2104
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2182..2303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2219..2238)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2273..2292
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2343..2478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2498..2517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2499..2517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2527..2544)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2527..2543)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2536..2558)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2685..3140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2772..2791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2982..3003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3166..3270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3271..3339,3340..4131)
                     /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(3489..3508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4302..4890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4791..4810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5044..5061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5178..5199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5214..5219,5220..5237,5238..5244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5252..5268
                     /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     5257..5279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5276..5292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5276..5292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5311..5331
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5313..5331
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5359..5585
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5586..5766
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5813..5938
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6431..6664
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6849..6893
                     /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        7068..7568
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7487..7505
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7580..7975
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8035..8152
                     /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 tagcatggcc gagttcccgt cgaaagttag cacgcggacc agcagtcctg
      601 cgcagggcgc cgaagcctcg gtgtcggcgc tgcgcccgga cctgggcttc gtgcgctccc
      661 gcctcggggc gctcatgctg ctgcagctgg tgctggggct gctggtgtgg gcgctgattg
      721 cggacacccc gtaccacctg tatccggcct atggctgggt gatgttcgtc gctgtcttcc
      781 tctggctggt gacaatcgtc ctcttcaacc tctacctgtt tcagctgcac atgaagttgt
      841 acatggttcc ctggccactg gtgttaatga tctttaacat cagcgccacc gttctctaca
      901 tcaccgcctt catcgcctgc tctgcggcag ttgacctgac atccctgagg ggcacccggc
      961 cttataacca gcgcgcggct gcctcgttct ttgcgtgttt ggtgatgatc gcctatggag
     1021 tgagtgcctt cttcagctac caggcctggc gaggagtagg cagcaatgcg gccaccagtc
     1081 agatggctgg cggctatgcc tacccaactt tcttgtacaa agtggttggt aagcctatcc
     1141 ctaaccctct cctcggtctc gattctacgt agtaatgagc tagcgctaac cggtggcgcg
     1201 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1261 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1321 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1381 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1441 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1501 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1561 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     1621 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     1681 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     1741 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     1801 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     1861 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     1921 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     1981 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2041 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2101 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2161 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     2221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2281 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     2341 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2401 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2461 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     2521 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2581 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2641 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2701 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2761 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2821 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2881 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2941 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3001 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3061 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3121 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3481 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3781 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4981 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5041 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5101 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5161 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5221 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5281 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     5341 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     5401 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     5461 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     5521 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     5581 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5641 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5701 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5761 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     5821 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     5881 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     5941 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     6001 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     6061 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     6121 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     6181 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     6241 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     6301 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     6361 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     6421 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     6481 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     6541 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     6601 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     6661 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     6721 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     6781 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     6841 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     6901 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     6961 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     7021 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccttg cgccttttcc
     7081 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     7141 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     7201 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     7261 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     7321 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     7381 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     7441 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     7501 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     7561 ctccccaggg ggtaccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     7621 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     7681 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     7741 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     7801 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     7861 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     7921 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggcctgca
     7981 gctgcagtag taagaattct agatcttgag acaaatggca gtattcatcc acaattttaa
     8041 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     8101 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     8161 ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg gttattaata
     8221 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
