LOCUS       sequence-319348-        7983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses FXYD3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 7983)
  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_FXYD3_OE_1
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        37..340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        341..544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     495..515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    567..591
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(853..877)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             879..920
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    962..1550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1015..1035)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1433..1444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1622..1855
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1933..2054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1970..1989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2024..2043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2094..2229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2249..2268)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2250..2268)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2278..2295)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2278..2294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2309)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2436..2891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2523..2542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2733..2754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2917..3021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3022..3090,3091..3882)
                     /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(3240..3259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4053..4641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4542..4561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4795..4812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4929..4950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4965..4970,4971..4988,4989..4995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5003..5019
                     /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     5008..5030
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5027..5043
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5027..5043
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5062..5082
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5064..5082
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5110..5336
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5337..5517
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5564..5689
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6182..6415
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6600..6644
                     /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        6819..7319
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7238..7256
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7331..7726
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7786..7903
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
       61 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      121 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      181 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      241 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      301 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      361 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      421 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      481 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      541 agctctctgg ctaactgtcg ggatcaacaa gtttgtacaa aaaagttggc atgcagaagg
      601 tgaccctggg cctgcttgtg ttcctggcag gctttcctgt cctggacgcc aatgacctag
      661 aagataaaaa cagtcctttc tactatgact ggcacagcct ccaggttggc gggctcatct
      721 gcgctggggt tctgtgcgcc atgggcatca tcatcgtcat gagtgcaaaa tgcaaatgca
      781 agtttggcca gaagtccggt caccatccag gggagactcc acctctcatc accccaggct
      841 cagcccaaag ctgcccaact ttcttgtaca aagtggttgg taagcctatc cctaaccctc
      901 tcctcggtct cgattctacg tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga
      961 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1021 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1081 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1141 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1201 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1261 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1321 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     1381 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     1441 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     1501 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     1561 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     1621 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     1681 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     1741 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     1801 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     1861 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     1921 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2041 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     2101 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2161 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2221 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     2281 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2341 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2401 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     2461 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2521 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2581 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2641 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2701 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2761 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2821 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2881 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3001 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3061 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3121 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3181 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3241 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3301 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3361 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3421 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3481 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3541 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3601 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3661 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3721 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3781 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3841 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3901 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3961 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4801 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4861 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4921 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4981 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5041 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     5101 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     5161 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     5221 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     5281 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     5341 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5401 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5461 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     5521 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     5581 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     5641 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     5701 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     5761 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     5821 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     5881 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     5941 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     6001 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     6061 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     6121 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     6181 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     6241 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     6301 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     6361 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     6421 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     6481 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     6541 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     6601 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     6661 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     6721 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     6781 tatcgtttca gacccacctc ccaaccccga ggggaccctt gcgccttttc caaggcagcc
     6841 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
     6901 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
     6961 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
     7021 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
     7081 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
     7141 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
     7201 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
     7261 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
     7321 gggtaccacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     7381 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     7441 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     7501 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     7561 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     7621 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     7681 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc agctgcagta
     7741 gtaagaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     7801 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     7861 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     7921 cagcagagat ccactttggc gccggctcga gggggcccgg ggttattaat agtaatcaat
     7981 tac
//
