LOCUS       sequence-334525-        8424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses HTR3D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8424)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8424)
  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_HTR3D_OE_1
FEATURES             Location/Qualifiers
     source          1..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    613..637
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      638..647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1340..1364)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1366..1407
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1449..2037
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1502..1522)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1920..1931)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2109..2342
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2420..2541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2457..2476)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2511..2530
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2581..2716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2736..2755)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2737..2755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2765..2782)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2765..2781)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2774..2796)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2923..3378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3220..3241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3404..3508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3509..3577,3578..4369)
                     /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(3727..3746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4540..5128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5029..5048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5282..5299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5416..5437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5452..5457,5458..5475,5476..5482)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5490..5506
                     /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     5495..5517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5514..5530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5514..5530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5549..5569
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5551..5569
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5597..5823
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5824..6004
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6051..6176
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6669..6902
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7087..7131
                     /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        7306..7806
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7725..7743
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7818..8213
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8273..8390
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggcgccggct cgagggggcc cggggttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      601 ctgtcgggat caacaagttt gtacaaaaaa gttggcacca tggtggccat caggcgcagg
      661 tgcaggccca gcccctacgt ggtaaacttt ctggtgccca gtggcattct gattgccatc
      721 gatgccctca gtttctacct gccactggaa agtgggaatt gtgccccatt caagatgact
      781 gttctgctgg gctacagcgt cttcctgctc atgatgaatg acttgctccc agccactagc
      841 acttcatcac atgcttcact agtacgtcct catccatcaa gagaccaaaa gcgaggtgtc
      901 tacttcgccc tgtgcctgtc cctgatggtg ggcagcctgc tggagaccat cttcatcacc
      961 cacctgctgc acgtggccac cacccagccc ctacctctgc ctcggtggct ccactccctg
     1021 ctgctgcact gcaccggcca agggagatgc tgtcccactg cgccccagaa gggaaataag
     1081 ggcccgggtc tcacccccac ccacctgccc ggtgtgaagg agccagaggt atcagcaggg
     1141 cagatgccag gccctgggga ggcagagctg acagggggct cagaatggac aagggcccag
     1201 cgggaacacg aggcccagaa gcagcactcg gtggagctgt gggtgcagtt cagccacgcg
     1261 atggacgccc tgctcttccg cctctacctg ctcttcatgg cctcctccat catcaccgtc
     1321 atatgcctct ggaacacctt gccaactttc ttgtacaaag tggttggtaa gcctatccct
     1381 aaccctctcc tcggtctcga ttctacgtag taatgagcta gcgctaaccg gtggcgcgtt
     1441 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1501 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1561 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     1621 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     1681 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     1741 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1801 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     1861 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     1921 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     1981 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     2041 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2101 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     2161 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     2221 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     2281 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     2341 cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     2401 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     2461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2521 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     2581 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2641 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2701 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     2761 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2821 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2881 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     2941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3061 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3361 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3481 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3541 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3601 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3661 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3721 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3781 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3841 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3901 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3961 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4021 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4081 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4141 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4201 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4261 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4321 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4381 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4441 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5161 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5221 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5281 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5341 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5401 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5461 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5521 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     5581 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     5641 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     5701 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     5761 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     5821 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5881 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5941 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6001 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     6061 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     6121 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     6181 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     6241 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     6301 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     6361 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     6421 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     6481 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     6541 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     6601 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     6661 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     6721 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     6781 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     6841 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     6901 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     6961 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7021 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7081 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7141 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7201 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7261 tcaccattat cgtttcagac ccacctccca accccgaggg gacccttgcg ccttttccaa
     7321 ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc
     7381 ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc
     7441 gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt
     7501 tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc
     7561 ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc
     7621 aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag
     7681 gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc
     7741 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
     7801 ccccaggggg taccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     7861 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     7921 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     7981 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     8041 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     8101 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     8161 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcctgcagc
     8221 tgcagtagta agaattctag atcttgagac aaatggcagt attcatccac aattttaaaa
     8281 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     8341 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     8401 acagggacag cagagatcca cttt
//
