LOCUS       sequence-319663-        8640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses PKD1L2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8640)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8640)
  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_PKD1L2_OE_1
FEATURES             Location/Qualifiers
     source          1..8640
                     /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(1474..1498)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1500..1541
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1583..2171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1636..1656)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2054..2065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2243..2476
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2554..2675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2591..2610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2645..2664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2715..2850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2870..2889)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2871..2889)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2899..2916)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2899..2915)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2908..2930)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3057..3512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3354..3375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3538..3642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3643..3711,3712..4503)
                     /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(3861..3880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4674..5262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5163..5182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5416..5433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5550..5571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5586..5591,5592..5609,5610..5616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5624..5640
                     /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     5629..5651
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5648..5664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5648..5664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5683..5703
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5685..5703
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5731..5957
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5958..6138
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6185..6310
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6803..7036
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7221..7265
                     /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        7440..7940
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7859..7877
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7952..8347
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8407..8524
                     /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 tggcatgggg gaggactctc cagtggctat gttcagctgg tatttggaca
      601 acaccccaac agagcaggct gagcccctcc cggatgcctg cagactcaga ggattttggc
      661 caaggtcctt aaccctcctc cagagcaaca cctccacgtt gctgttgaac agctcgtttc
      721 tgcagtcccg gggagaggtc atccgaatca gagccacagc actgaccagg catgcctatg
      781 gggaggacac ctatgtgatc agcactgtgc ctccccgtga ggtgcctgcc tgcactattg
      841 ccccagagga gggcaccgtt ctgacgagct ttgccatctt ctgcaacgcc tccacagccc
      901 tgggacccct ggagttctgc ttctgtctgg aatcaggttc ctgcctacac tgtggccctg
      961 aacctgccct cccatcagtg tatctgccac ttggagagga gaacaatgac tttgtgctga
     1021 cagtagttat ttctgccacc aatcgtgcag gggacacgca gcagacccag gccatggcta
     1081 aggtggcact cggagacaca tgtgttgagg atgtagcatt ccaggctgcc gtgtcagaga
     1141 aaatccccac agctctgcaa ggcgagggtg gccccgagca gctcctccag ctggccaagg
     1201 ctgtgtcctc catgctgaac caagagcatg aaagccaggg ctcaggacag tcactgagca
     1261 tagacgtcag acagaaggtc agagagcatg tgctgggatc actgtctgca gtcaccaccg
     1321 gcttggagga cgtgcagagg gtgcaggagc tggccgaggt gctgagagag gtgacctgcc
     1381 ggagtaagga actcacaccc tcggcccagg ggtcctgcat gggcgattca tgggaaggtg
     1441 cccctcctgc tgcccatgta tctcacgcta ggtgcccaac tttcttgtac aaagtggttg
     1501 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gtagtaatga gctagcgcta
     1561 accggtggcg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     1621 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1681 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1741 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     1801 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     1861 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     1921 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     1981 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2041 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2101 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2161 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     2221 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     2281 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     2341 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     2401 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     2461 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     2521 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     2581 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2641 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     2701 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2761 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2821 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     2881 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2941 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     3061 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3121 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3181 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3241 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3301 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3361 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3421 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3481 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     3541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4201 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4501 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5341 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5401 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     5461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5521 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     5581 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     5641 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     5701 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     5761 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     5821 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     5881 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     5941 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6001 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6061 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6121 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     6181 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     6241 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     6301 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     6361 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     6421 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     6481 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     6541 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     6601 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     6661 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     6721 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     6781 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     6841 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     6901 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     6961 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     7021 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     7081 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     7141 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     7201 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     7261 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     7321 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     7381 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccct
     7441 tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt
     7501 ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg
     7561 tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc
     7621 taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac
     7681 gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc
     7741 gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag
     7801 gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt
     7861 gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa
     7921 tcaccgacct ctctccccag ggggtaccac catggccaag cctttgtctc aagaagaatc
     7981 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     8041 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     8101 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     8161 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     8221 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     8281 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     8341 ggagggcctg cagctgcagt agtaagaatt ctagatcttg agacaaatgg cagtattcat
     8401 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     8461 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     8521 ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg agggggcccg
     8581 gggttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
